Litchi extracting solution and application of litchi extracting solution as oil control component in skin oil balancing effect judgment method
By using lychee extract to regulate multiple skin regulatory factors, the problem of unstable evaluation of existing oil-control products has been solved, achieving skin oil balance and alleviating problems, and providing an accurate method for judging the effect.
Patent Information
- Application Number
- CN202511060611.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-30
- Publication Date
- 2025-11-25
AI Technical Summary
Existing oil-control products ignore multiple regulatory factors when verifying their oil-control effects, resulting in unstable evaluations, an inability to effectively balance skin oil, and a lack of a comprehensive judgment method that considers multiple regulatory pathways.
Using lychee extract as an oil-controlling component, it contains polyphenols and flavonoids, which regulate the expression of SRD5A2, FGFR1, AKT1 and ACLY genes in the skin, reduce the expression of SESN2 gene, and increase the expression of SESN2 gene. Combined with butylene glycol, it improves skin compatibility, inhibits excessive sebum secretion, and balances skin oil.
It achieves effective balance of skin oil, alleviates skin problems such as blackheads, acne and enlarged pores, and provides a simple and accurate method to judge the effect of balancing skin oil.
Smart Images
Figure CN121003573A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of skin oil control, and particularly relates to a method for judging the effect of balancing skin oil by using litchi extract as an oil control component. BACKGROUND
[0002] The content and components of sebum have a significant impact on the skin condition, and excessive secretion of skin oil can lead to the formation of acne, acne and enlarged pores. Existing oil control products all emphasize the realization of oil control effect, but excessive inhibition of skin oil secretion breaks the dynamic balance of skin oil secretion, and instead promotes the sebaceous glands to secrete more oil, leading to skin secretion metabolic disorder and damaged skin barrier function.
[0003] Currently, the verification of oil control effect in the field of cosmetics mainly adopts a verification scheme of 5alpha reductase inhibition model. 5alpha reductase, as one of the regulatory factors of human oil production, plays a key role in the process of converting testosterone into dihydrotestosterone, and dihydrotestosterone has an important regulating function on the development and secretion of sebaceous glands. In theory, inhibiting the activity of 5alpha reductase can reduce oil secretion. However, 5alpha reductase is only one of the many regulatory factors of human oil production, and this model ignores other regulatory factors and has a single regulating pathway. The oil control effect of the composition evaluated is not stable, or shows excessive inhibition of oil, or has poor oil control effect, and cannot achieve the expected effect. At present, the oil control path of the oil control product is single, and the oil control effect of the oil control product lacks an effective judgment method that can comprehensively consider multiple regulating pathways, multiple regulating factors and simulate the real physiological environment of the skin. SUMMARY
[0004] The present application provides a litchi extract with the effect of balancing skin oil.
[0005] Another object of the present application is to provide the litchi extract for use as an oil control component in cosmetics to balance skin oil.
[0006] Another object of the present application is to provide a method for testing the effect of balancing skin oil, which tests the skin after using cosmetics containing an oil control component, and can test whether the cosmetics containing the oil control component have the effect of balancing skin oil.
[0007] To solve the above technical problems, the technical solution provided by the present application is: A litchi extract contains the following components by mass fraction: litchi fruit extract 2-12 parts, butanediol 35-55 parts.
[0008] Preferably, the total flavonoids in the litchi extract are ≥2000ppm, the total phenol is ≥2000ppm, and the total sugar is ≤5%.
[0009] The litchi extract of the present invention also contains the following components in parts by weight: 10-20 parts glycerol and 13-53 parts water.
[0010] The litchi extract provided by this invention contains polyphenols and flavonoids, which can both reduce the expression of skin regulatory factors SRD5A2, FGFR1, AKT1 and ACLY genes and increase the expression of skin regulatory factor SESN2 gene. Butylene glycol can promote the compatibility of litchi active substances with skin sebum components. When the litchi extract obtained by combining the two is applied to the skin, it can inhibit excessive secretion of sebaceous glands, balance skin oil, and has a beautifying effect.
[0011] The present invention also provides the lychee extract as an oil-controlling component for use in cosmetics to balance skin oil.
[0012] Lychee extract is rich in polyphenols, flavonoids and other active ingredients that can inhibit excessive sebum secretion. This invention applies lychee extract to cosmetics as an oil-controlling component, which can effectively balance skin oil and thus alleviate skin problems such as acne, pimples, enlarged pores and oily sensitive skin.
[0013] This invention also provides a method for testing the effect of balancing skin oil. The method involves testing skin after using a cosmetic containing oil-controlling ingredients. If either condition B or C is not met, the oil-controlling ingredient is determined to have no effect on balancing skin oil. Conditions B and C are specifically as follows: B. The expression levels of regulatory factors SRD5A2, FGFR1, AKT1, and ACLY genes in the skin were significantly lower than those in the negative control group. C. The expression level of the regulatory factor SESN2 gene in the skin was significantly higher than that in the negative control group.
[0014] 1. The method for judging the effect of the skin oil control scheme on oil balance provided by the present invention uses oil control components to test the oil control effect on the skin. The actual effect on skin oil balance is judged by detecting whether the oil control components meet both condition B, which regulates the expression levels of SRD5A2, FGFR1, AKT1 and ACLY genes to be lower than the negative control group, and condition C, which regulates the expression level of SESN2 gene to be higher than the negative control group. The principle is detailed as follows, where (1)~(4) are condition B and (5) is condition C: (1) The SRD5A2 gene regulates sebum secretion by regulating androgen metabolism. Its gene expression level is lower than that of the negative control group. It inhibits the conversion of androgens (such as testosterone) into more active dihydrotestosterone (DHT), thereby inhibiting the binding of DHT to AR in sebaceous gland cells. Downstream signaling pathways (such as insulin resistance PI3K / AKT1 signaling pathway, steroid biosynthesis, etc.) are blocked, thereby inhibiting excessive sebum secretion.
[0015] (2) FGFR1 is an important regulator of sebaceous gland development and lipid metabolism. Its gene expression level is lower than that of the negative control group. It can effectively weaken the signal transduction of FGF (such as FGF2 and FGF9) in the skin, thereby resulting in insufficient activation of downstream signaling pathways (such as MAPK and PI3K-AKT1 pathways), and ultimately inhibiting excessive secretion of sebum.
[0016] (3) AKT1 is the core regulator of the PI3K-AKT-mTOR signaling pathway. Its gene expression level is lower than that of the negative control group, which can reduce mTORC1 activity. Reduced mTORC1 activity can reduce the cleavage and nuclear translocation of SREBP (sterol regulatory element binding protein), thereby downregulating the expression of key lipid synthesis enzymes (such as FASN, SCD1, HMG-CoA), and ultimately inhibiting excessive lipid secretion.
[0017] (4) ACLY is a key enzyme that catalyzes the breakdown of citric acid into acetyl-CoA and oxaloacetate. Its gene expression level is lower than that of the negative control group, which can significantly reduce the level of acetyl-CoA in the cytoplasm. Since the basic steps of fatty acid synthesis (such as malonyl-CoA generation and fatty acid chain elongation) depend on acetyl-CoA, it ultimately inhibits excessive secretion of lipids.
[0018] (5) SESN2 can mediate the oxidative stress process of cells. Its gene expression level is higher than that of the negative control group. It can inhibit the transcriptional activity of PPAR-γ, reduce the expression of its target genes (such as lipoprotein lipase LPL and fatty acid transporter FATP), reduce the breakdown and uptake of fatty acids, thereby inhibiting excessive secretion of lipids.
[0019] 2. The judgment method provided by the present invention is simple to operate and accurate in results. This judgment method can be used to test the oil-balancing effect of products, and can be used to test various skin oil control solutions. It can judge whether various skin oil control solutions have the oil-balancing effect, and screen out skin oil control solutions with the oil-balancing effect. It can also screen oil control composition formulas with the oil-balancing effect.
[0020] The present invention also provides a method for preparing the litchi extract, comprising the following steps: S1. The litchi fruit was crushed and extracted by ultrasonic reflux to obtain crude litchi fruit extract; S2. The crude extract of litchi fruit is subjected to molecular imprinting adsorption to obtain litchi fruit extract; S3. Glycerin, butylene glycol, and water were added to the litchi fruit extract in proportion to obtain a total flavonoid concentration of 4000-6000 ppm and a total phenol concentration of 4000-6000 ppm, thus obtaining litchi extract concentrate 1. S4. The concentrated litchi extract 1 was subjected to ultrafiltration and nanofiltration to obtain the retentate; S5. The retentate is post-processed to obtain the litchi extract.
[0021] Preferably, the method for judging the effect of balancing skin oil includes the following steps: S1. Cell viability test; S2. The expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2 genes in the sample group, blank group, control group, and negative control group were tested. The expression levels of each gene obtained from the test were used to determine whether the skin oil control program had a balancing effect on oil production.
[0022] Preferably, the cells in S1 are human sebaceous gland cells SZ95.
[0023] Preferably, S1 specifically comprises: S11. After culturing human sebaceous gland cells, the same volume of test samples of different concentrations were added to obtain human sebaceous gland cells X1; S12. Add MTT solution to human sebaceous gland cells X1 to obtain human sebaceous gland cells X2; S13. Discard the liquid in human sebaceous gland cells X2, add DMSO, and measure the absorbance using an ELISA reader.
[0024] Preferably, the method for testing regulatory factors in S2 is transcriptome sequencing RNA-seq.
[0025] More preferably, the specific steps of transcriptome sequencing RNA-seq are as follows: S21. Human sebaceous gland cells SZ95 were divided into sample group, blank group, control group and negative control group and cultured separately. S22. RNA extraction: The human sebaceous gland cells SZ95 obtained in S21 were lysed, genomic DNA was removed, RNA was adsorbed, impurities were removed, RNA was eluted, and RNA purity and concentration were detected. S23. cDNA Synthesis: Enrich the mRNA extracted from S22 and reverse transcribe the mRNA into cDNA; S24. Adapter ligation: cDNA is ligated to a specific adapter sequence and an index is added to obtain the ligation product; S25. Using the ligation product as a template, amplify to obtain a cDNA library; The S26.cDNA library was sequenced using high-throughput sequencing to calculate the expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2, and to determine whether each gene was higher or lower than the negative control group.
[0026] The litchi variety in the litchi extract of this invention is selected from one or more of the following varieties: Fei Zi Xiao, Gui Wei, Nuo Mi Ci, Xian Jin, and Gua Lü. Attached Figure Description
[0027] Figure 1 This is a fluorescence photograph obtained from the blank control group.
[0028] Figure 2 This is a fluorescence photograph obtained from the negative control group.
[0029] Figure 3 This is a fluorescence photograph obtained from the positive control group.
[0030] Figure 4 The fluorescence image is obtained by adding 0.1% oil-controlling component 1. Detailed Implementation
[0031] The present invention will be further described in detail below with reference to specific embodiments.
[0032] Example 1 I. Preparation of litchi extract as an oil-controlling component Oil-controlling component 1 A lychee extract contains the following components in parts by weight: 8 parts lychee fruit extract, 40 parts butylene glycol, 15 parts glycerol, and 30 parts water; the lychee extract contains 2000-3000 ppm total flavonoids, 2000-3000 ppm total phenols, and ≤5% total sugar.
[0033] The method for preparing litchi extract includes the following steps: S1. First, crush fresh Guiwei lychee fruit into pieces with a diameter of ≤1cm, add 4~6 times the amount of pure water; then, after ultrasonic reflux extraction for 1 hour, filter it through a 0.45 micron filter membrane until clear, and concentrate the filtrate to 400% of the raw drug amount by 150~300D nanofiltration to obtain crude lychee fruit extract. S2. Adsorption and separation of molecularly imprinted polymer: First, the loading ratio is the volume of the concentrate, and the amount of molecularly imprinted polymer is 4 mL / g. The mixture is ultrasonicated for 0.5 h with 60-80% butanediol (4 times the mass of the molecularly imprinted polymer), filtered to obtain a filtrate, and then concentrated to 300% of the crude drug amount using 150-300D nanofiltration. Then, the solvent is added and ultrasonicated twice more. The ultrasonic eluates are combined to finally obtain the litchi fruit extract. The molecularly imprinted polymer is the one obtained in Example 2 of patent CN 2024114979016. S3. Glycerin, butylene glycol, and water were added to litchi extract concentrate 1 in a certain proportion until the total flavonoids reached 4000-6000 ppm and the total phenols reached 4000-6000 ppm, thus obtaining litchi extract concentrate 1; the total flavonoid content was detected by the sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method using an ultraviolet spectrophotometer; the total phenol content was detected by method 2 of GB / T 8313-2008; S4. The concentrated litchi extract 1 was ultrafiltered at 10000D to obtain the permeate. The permeate was then nanofiltered at 150~300D to obtain the retentate. The retentate contained 2000~3000ppm total flavonoids, 2000~3000ppm total phenols, and ≤5% total sugar. The total flavonoid content was determined by the sodium nitrite-aluminum nitrate-sodium hydroxide colorimetric method using a UV spectrophotometer. The total phenol content was determined by Method II of GBT 8313-2008. The total sugar content was determined by the phenol-sulfuric acid colorimetric method using a UV spectrophotometer. S5. The retentate is then post-treated as follows: after being kept at 90~95℃ for 0.5h, the retentate is filtered through a 0.22μm membrane to obtain the litchi extract.
[0034] Oil-controlling component 2 A lychee extract, which differs from oil-controlling component 1 in that it contains the following components by weight: 2 parts lychee fruit extract, 35 parts butylene glycol, 10 parts glycerin, and 13 parts water; wherein the lychee variety of the lychee fruit extract is Feizixiao and Xianjinfeng in equal weight proportions.
[0035] Oil-controlling component 3 A lychee extract, which differs from oil-controlling component 1 in that it contains the following components by weight: 12 parts lychee fruit extract, 55 parts butylene glycol, 20 parts glycerin, and 53 parts water; wherein the lychee variety of the lychee fruit extract is Nuomici and Guailv in equal weight proportions.
[0036] Oil-controlling component 4 A litchi extract, which differs from oil-controlling component 1 in that it contains the following components by weight: 1 part litchi fruit extract, 30 parts butylene glycol, 5 parts glycerin, and 8 parts water; the litchi extract contains 1000~1500 ppm of total flavonoids.
[0037] Oil-controlling component 5 A litchi extract, which differs from oil-controlling component 1 in that it contains the following components by weight: 15 parts litchi fruit extract, 60 parts butylene glycol, 25 parts glycerin, and 60 parts water; the litchi extract contains 1000~1500 ppm total phenols.
[0038] 2. Assess the skin after using the oil-controlling ingredients. If the skin does not meet either of the following conditions B or C, then the oil-controlling ingredients are deemed not to have the effect of balancing skin oil. Conditions B and C are specifically as follows: B. The expression levels of human skin regulatory factors SRD5A2, FGFR1, AKT1 and ACLY genes were significantly lower than those in the negative control group. C. The expression level of the regulatory factor SESN2 gene in human skin was significantly higher than that in the negative control group.
[0039] 1. Cell viability assay (MTT) The cell viability test steps and results for oil-controlling component 1 are as follows: S11. First, human sebaceous gland cell suspension was seeded into a 96-well cell culture plate and cultured for 24 hours; then, oil-controlling component 1 was taken, dissolved in the culture medium, and fully dissolved to obtain the test sample; finally, the same volume of test sample with different concentrations was added to each well, and the cells were returned to the incubator for incubation to obtain human sebaceous gland cells X1. S12. Add MTT solution to each well of human sebaceous gland cells X1 and incubate in an incubator to obtain human sebaceous gland cells X2; S13. Discard the liquid in human sebaceous gland cells X2, add DMSO, and measure the absorbance using an ELISA reader.
[0040] The cell viability testing methods for oil-controlling components 2-5 were the same as those for oil-controlling component 1, with the cell viability test results for oil-controlling component 5 shown in Table 2. As can be seen from Tables 1 and 2, when the concentration of oil-controlling component 1 or oil-controlling component 5 was ≤0.1%, the cell viability was greater than 90%, and both could be used for subsequent experiments. The cell viability detection for oil-controlling components 2-4 was similar to that for oil-controlling components 1 and 5; when the sample concentration (oil-controlling components 2-4) was ≤0.1%, the cell viability was greater than 90%, and all could be used for subsequent experiments.
[0041] Assuming the cell viability of the negative control group is 100%, the relative cell viability of each group is calculated using the following formula.
[0042]
[0043] OD TA The three-fold average of the optical density of the tested sample group. OD c The average of three optical density measurements in the negative control group. OD B The average of the optical density of the blank control group was measured three times.
[0044] Table 1. Relative cell activity at different concentrations of oil-controlling component 1
[0045] Table 2. Relative cell activity at different concentrations of oil-controlling component 5.
[0046] 2. The expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2 in human sebaceous gland cells SZ95 were detected using transcriptome sequencing RNA-seq, and the upregulation / downregulation of each regulatory factor relative to the negative control group was determined (judgment criterion: Fold Change). The transcriptome sequencing RNA-seq method is as follows: S21. Human sebaceous gland cells SZ95 were divided into sample groups (each sample group was supplemented with 0.1% oil-controlling component 1-5), blank control group, and negative control group, and cultured separately. S22. RNA extraction: The human sebaceous gland cells SZ95 obtained in S21 were lysed, genomic DNA was removed, RNA was adsorbed, impurities were removed, RNA was eluted, and RNA purity and concentration were detected. S23. cDNA Synthesis: Enrich the mRNA extracted from S22 and reverse transcribe the mRNA into cDNA; S24. Adapter ligation: cDNA is ligated to a specific adapter sequence and an index is added to obtain the ligation product; S25. Library amplification: Using the ligation product as a template, amplify to obtain a cDNA library; The S26.cDNA library was subjected to high-throughput sequencing to calculate the expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2, and to determine whether each gene was higher or lower than the negative control group. The specific steps included: (1) High-throughput sequencing: The mixed cDNA library is loaded onto the Illumina sequencing platform, and the sequencer performs paired-end sequencing on the cDNA fragments in the library, generating a large number of short-read raw reads; (2) Data quality control and preprocessing: First, the raw reads obtained from sequencing were subjected to quality control. The software FastQC was used to detect the base quality, GC content, adapter contamination, etc. of the raw data. Then, tools such as Trimmomatic were used to remove low-quality reads, adapter sequences, and contaminating sequences to obtain high-quality clean reads. (3) Sequence alignment: Align clean reads with the reference genome or reference transcriptome using HISAT2; through alignment, determine the position of each read in the genome or transcriptome and identify which reads come from the target gene domain; (4) Gene expression quantification: Count the number of reads aligned to each gene region to obtain the raw count of the gene, i.e., how many reads come from the target gene; (5) Differential expression analysis: Statistical modeling and inference based on the negative binomial distribution were performed using the DESeq2 R package to identify differentially expressed genes (DEGs) in the treatment group relative to the negative control group. The core computational steps included: (a): Calculating the size factor (purpose: to correct for differences in sequencing depth between samples): s_j = median(gene count_ij / geometric mean count_i), s_j: Size factor of sample j.
[0047] Gene count_ij: The original count of gene i in sample j. Geometric mean count_i: The geometric mean of the counts of gene i across all samples. Median: Calculate the gene count _ij / geometric mean count _i for all genes, and then take the median of these ratios as s_j for sample j.
[0048] (b): Calculate the standardized within-group mean expression level: Within-group mean expression level = mean (gene count_ij / s_j) (c): Calculate Log2 Fold Change (log2FC): log2FC_i = log2(normalized mean expression level of gene i in the treatment group / normalized mean expression level of gene i in the control group) (d): Calculate the p-value of the target gene using the Wald test. (6) Differentially expressed gene screening and Fold Change calculation: First, candidate genes were screened based on statistical significance: genes with a P value < 0.05 were selected. This invention screened 36 genes with significant differences, including SRD5A2, FGFR1, AKT1, ACLY, SESN2, SLC27A1, and AGPAT.
[0049] Secondly, calculate the Fold Change (FC) of these significantly differentially expressed genes: Fold Change = 2^(log2FC) (7) Determine whether the SRD5A2, FGFR1, AKT1, ACLY and SESN2 genes are upregulated or downregulated relative to the negative control group. The criteria are: Fold Change>1, the gene expression level is higher than that of the negative control group, and the gene is upregulated; Fold Change<1, the gene expression level is lower than that of the negative control group, and the gene is downregulated.
[0050] The gene expression levels of each regulatory factor sample group and the gene expression levels of the blank control group were calculated and compared with the negative control group. The calculation results are shown in Table 3.
[0051] Table 3. Ratio of regulatory factor gene expression levels in human sebaceous gland cells SZ95 compared to the negative control group
[0052] Fold Change 1: (1) The fold increase or decrease in gene expression compared to the negative control group (NC); "Fold change" > 1 indicates that the gene expression level is increased, that is, the gene expression level is higher than that of the negative control group; "Fold change" < 1 indicates that the gene expression level is decreased, that is, the gene expression level is higher than that of the negative control group.
[0053] (2) In the RNA-seq method detection, the S26.cDNA library was sequenced by high-throughput sequencing. (5) Differential expression analysis has been performed by Wald test to calculate the p value of the target gene, and candidate genes were screened based on statistical significance: genes with p value <0.05 were selected. In this invention, 36 genes with significant differences were screened, including SRD5A2, FGFR1, AKT1, ACLY, SESN2, SLC27A1 and AGPAT. Therefore, the obtained Fold Change value also has significant differences.
[0054] Table 3 shows that oil-controlling components 1-3 meet both conditions B and C and have an oil-controlling effect; oil-controlling component 4 does not meet conditions B and C, and oil-controlling component 5 does not meet condition B, so neither of them has the effect of balancing skin oil.
[0055] III. Human Sebum Secretion Inhibition Rate Detection Human sebaceous gland cells SZ95 were used as model cells. The cells were stained with Nile Red, a lipophilic fluorescent dye, and photographed under a fluorescence microscope. The collected images were analyzed for fluorescence intensity using ImageJ software to calculate the sebum secretion inhibition rate. The specific detection method is as follows: 1. Cell viability test (MTT): The results are similar to those in step 3. When the sample concentration is ≤0.1%, the cell viability is greater than 90%, and both can be used for subsequent experiments.
[0056] 2. Fluorescence intensity test of sebum secretion (1) Routine cell culture. Human sebaceous gland cell suspension SZ95 was seeded into 96-well cell culture plates and cultured for 24 hours; (2) Each group should add the corresponding substances according to Table 4 and incubate for 24 hours; (3) After the culture is completed, wash with phosphate buffer solution and add paraformaldehyde, then let stand at room temperature; (4) Add Triton X-100 and let stand at room temperature; (5) Add Nile Red working solution and incubate at room temperature in the dark; (6) Take photos under a fluorescence microscope while avoiding light; among which: Figure 1 These are fluorescence images obtained from the blank control group. Figure 2 These are fluorescence images obtained from the negative control group. Figure 3 These are fluorescence images obtained from the positive control group. Figure 4 The fluorescence image is obtained by adding 0.1% oil-controlling component 1.
[0057] (7) The collected photos were analyzed using ImageJ software. The lipid secretion inhibition rate was calculated according to the following formula. The calculation results are shown in Table 5.
[0058]
[0059] Table 4. Reagents added to each experimental group in the sebum secretion inhibition test.
[0060] Table 5. Human sebum secretion inhibition rate
[0061] “ns” indicates no significant difference (P ≥ 0.05); * indicates a statistically significant difference compared to the negative control group (0.01 ≤ P < 0.05); ** indicates an extremely significant difference compared to the negative control group (P < 0.01). Figures 1-4 In the diagram, red fluorescence represents lipid droplets; the stronger and more obvious the red fluorescence, the more sebum is secreted. Figure 1 As can be seen, without the addition of oleic acid for induction, the red fluorescence intensity is weak, and the cells secrete a small amount of lipid; Figure 2 It can be seen that the addition of oleic acid induces an increase in the intensity of red fluorescence, and the cells secrete a large amount of lipids; Figure 3 It can be seen that the intensity of red fluorescence is significantly reduced when positive substances are added compared with the negative control group; Figure 4 It can be seen that the addition of 0.1% oil-controlling component 1 significantly reduced the intensity of red fluorescence.
[0062] Combination Figures 1-4 As can be seen from Table 5, oil-controlling component 1 has the effect of balancing skin oil, while oil-controlling components 4 and 5 do not have the effect of balancing skin oil, which is consistent with the conclusion of the judgment method provided by the present invention.
[0063] Comparative Example 1 To verify the effectiveness of this evaluation method, another skin oil control method was selected to evaluate its oil-balancing effect. This method selects regulatory factors on the skin that may have an oil-balancing effect. Compared with Example 1, the same method was used to evaluate the oil-balancing effect by comparing the expression levels of SRD5A2, ACLY, and SESN2 genes with those higher / lower than the negative control group. The difference was that SLC27A1 and AGPAT replaced FGFR1 and AKT1. Theoretically, reducing the expression level of SLC27A1 protein can enhance fatty acid transport and inhibit lipid accumulation; reducing the expression level of AGPAT protein can regulate triglyceride biosynthesis; and increasing the expression level of SESN2 protein can activate the AMPK pathway, increase the production of sex hormone-binding globulin SHBG, and promote sebum secretion. Theoretically, these five factors can play a role in balancing oil. To verify whether it can actually play a role in balancing oil, the oil-balancing effects of oil-controlling components 4 and 5 were evaluated using three targets: SRD5A2, ACLY, SESN2, SLC27A1, and AGPAT. Specifically: 1. Cell viability assay (MTT) was performed in Example 1. When the sample concentration was ≤0.1%, the cell viability was greater than 90%, and both could be used for subsequent experiments.
[0064] 2. The expression levels of SLC27A1 and AGPAT genes, regulatory factors in human sebaceous gland cells SZ95, were detected using transcriptome sequencing RNA-seq.
[0065] The specific detection method is the same as in Example 1. The gene expression levels of SLC27A1 and AGPAT sample groups and the gene expression levels of the blank control group were calculated as ratios to the negative control group. The calculation results are shown in Table 6.
[0066] Table 6. Ratio of regulatory factor gene expression levels in human sebaceous gland cells SZ95 to the negative control group
[0067] Fold Change 1: (1) The fold increase or decrease in gene expression compared to the negative control group (NC); "Fold change" > 1 indicates that the gene expression level is increased, that is, the gene expression level is higher than that of the negative control group; "Fold change" < 1 indicates that the gene expression level is decreased, that is, the gene expression level is higher than that of the negative control group.
[0068] (2) In the RNA-seq method detection, the differential expression analysis of the S26.cDNA library after high-throughput sequencing in step (5) has been calculated by Wald test to calculate the p value of the target gene, and candidate genes are screened based on statistical significance: genes with p value <0.05 are selected. In this invention, 36 genes with significant differences were screened, including SRD5A2, FGFR1, AKT1, ACLY, SESN2, SLC27A1 and AGPAT. Therefore, the obtained Fold Change value is also significantly different.
[0069] Table 6 shows that oil-controlling components 4 and 5 meet the conditions after gene replacement and have the effect of balancing skin oil. However, they are contrary to the results of the human sebum secretion inhibition rate test. Therefore, the judgment method obtained by replacing FGFR1 and AKT1 with SLC27A1 and AGPAT cannot accurately determine the effect of oil-controlling components on balancing skin oil.
[0070] The above description is merely an embodiment of the present invention and does not limit the scope of patent protection. Any non-substantial changes or substitutions made by those skilled in the art based on the present invention will still fall within the scope of patent protection.
Claims
1. A litchi extract, characterized in that, It contains the following components by weight: 2-12 parts of lychee fruit extract and 35-55 parts of butylene glycol.
2. The litchi extract according to claim 1, characterized in that, The litchi extract contained ≥2000ppm total flavonoids, ≥2000ppm total phenols, and ≤5% total sugar.
3. The method for preparing the litchi extract according to any one of claims 1 to 2, characterized in that, Includes the following steps: S1. The litchi fruit was crushed and extracted by ultrasonic reflux to obtain crude litchi fruit extract; S2. The crude extract of litchi fruit is subjected to molecular imprinting adsorption to obtain litchi fruit extract; S3. Glycerin, butylene glycol, and water were added to the litchi fruit extract in proportion to obtain a total flavonoid concentration of 4000-6000 ppm and a total phenol concentration of 4000-6000 ppm, thus obtaining litchi extract concentrate 1. S4. The concentrated litchi extract 1 was subjected to ultrafiltration and nanofiltration to obtain the retentate; S5. The retentate is post-processed to obtain the litchi extract.
4. The lychee extract according to any one of claims 1 to 2 is used as an oil-controlling component in cosmetics to balance skin oil.
5. A method for judging the effect of balancing skin oil, characterized in that, Skin samples were tested after using cosmetics containing oil-controlling ingredients. If either condition B or C was not met, the oil-controlling ingredient was deemed not to have a balancing effect on skin oil. Conditions B and C are as follows: B. The expression levels of regulatory factors SRD5A2, FGFR1, AKT1, and ACLY genes in the skin were significantly lower than those in the negative control group. C. The expression level of the regulatory factor SESN2 gene in the skin was higher than that in the negative control group and the difference was statistically significant.
6. The method for judging the effect of balancing skin oil according to claim 5, characterized in that, The following steps are included: S1. Cell viability test; S2. The expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2 genes in the sample group, blank group, control group, and negative control group were tested. The expression levels of each gene obtained from the test were used to determine whether the skin oil control program had a balancing effect on oil production.
7. The method for judging the effect of balancing skin oil according to claim 6, characterized in that, The cells in S1 are human sebaceous gland cells SZ95.
8. The method for judging the effect of balancing skin oil according to claim 6, characterized in that, Specifically, S1 is: S11. After culturing human sebaceous gland cells, the same volume of test samples of different concentrations were added to obtain human sebaceous gland cells X1; S12. Add MTT solution to human sebaceous gland cells X1 to obtain human sebaceous gland cells X2; S13. Discard the liquid in human sebaceous gland cells X2, add DMSO, and measure the absorbance using an ELISA reader.
9. The method for judging the effect of balancing skin oil according to claim 6, characterized in that, The expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2 in S2 were measured using transcriptome sequencing RNA-seq.
10. The method for judging the effect of balancing skin oil according to claim 9, characterized in that, The specific steps of transcriptome sequencing RNA-seq are as follows: S21. Human sebaceous gland cells SZ95 were divided into sample group, blank group, control group and negative control group and cultured separately. S22. RNA extraction: The human sebaceous gland cells SZ95 obtained in S21 were lysed, genomic DNA was removed, RNA was adsorbed, impurities were removed, RNA was eluted, and RNA purity and concentration were detected. S23. cDNA Synthesis: Enrich the mRNA extracted from S22 and reverse transcribe the mRNA into cDNA; S24. Adapter ligation: cDNA is ligated to a specific adapter sequence and an index is added to obtain the ligation product; S25. Using the ligation product as a template, amplify to obtain a cDNA library; The S26.cDNA library was sequenced using high-throughput sequencing to calculate the expression levels of regulatory factors SRD5A2, FGFR1, AKT1, ACLY, and SESN2, and to determine whether each gene was higher or lower than the negative control group.