Application of querceric acid

Abstract

The invention discloses application of querceric acid. The querceric acid can be applied to the field of cosmetics as an active ingredient to achieve the effects of inhibiting sebaceous gland secretion and controlling skin oil. The invention has the advantage of skin oil control when applied to cosmetics.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to the application of quercetin. Background Technology

[0002] Clinically, based on the amount of lipids on the skin's surface, skin is broadly classified into several types, including oily skin, combination skin, normal skin, and dry skin. Oily skin is prone to greasiness and enlarged pores, affecting appearance and increasing the risk of related skin conditions such as acne, pimples, and seborrheic dermatitis. Research has found that oily skin is caused by excessive sebum secretion from the sebaceous glands. Therefore, inhibiting excessive sebum secretion is key to controlling oil production and maintaining healthy skin.

[0003] Sebaceous glands are mostly located between hair follicles and arrector pili muscles. They consist of one or more sac-like acini connected by a common short duct, and are most abundant on the forehead, nose, and upper back. They function to lubricate the skin and hair, regulate body temperature, and have antibacterial properties. When sebaceous gland cells divide and migrate, they form small lipid droplets. When the gland cells disintegrate, these droplets are expelled together to form sebum. Furthermore, sebaceous gland cells express various hormone receptors, and the development and secretion of sebaceous glands are strongly regulated by sex hormones. If sebum cannot be expelled in time, it can easily lead to blockage of the sebaceous gland duct, causing bacterial infection and triggering acne.

[0004] Roburic acid (12-dien-3-oic acid), also known as quercetin, is a natural tetracyclic triterpenoid compound. Figure 1 As shown, it was first discovered in oak galls, hence its name. Quercetin has strong biological activities in antioxidation, anti-inflammation, anti-cancer, and anti-osteoarthritis.

[0005] Currently, quercetin is mainly used in anti-inflammatory, anti-cancer, and antioxidant fields, but no research has been found on its application in oil control. Summary of the Invention

[0006] The purpose of this invention is to provide an application of quercetin. This invention has the advantage of achieving skin oil control effects when applied in cosmetics.

[0007] The technical solution of the present invention: The application of quercetin as an active ingredient in cosmetics with skin oil-controlling effects.

[0008] In the aforementioned application of quercetin, the mass percentage of quercetin added to the cosmetic is 0.000025%-0.0004%.

[0009] In the aforementioned application of quercetin, the mass percentage of quercetin added to the cosmetic is 0.0002%.

[0010] Compared with the prior art, the beneficial effects of this application are as follows: This invention provides an application of quercetin, which can be used alone as an active ingredient in the field of cosmetics to achieve the effects of inhibiting sebaceous gland secretion and controlling skin oil.

[0011] Therefore, this invention has the advantage of achieving skin oil control when applied in cosmetics. Attached Figure Description

[0012] Figure 1 It is the structural formula of quercetin; Figure 2 This is a graph showing the lipid accumulation detection in human sebaceous gland cells during Experiment 2. Detailed Implementation

[0013] The present invention will be further described below with reference to the accompanying drawings and embodiments, but this should not be construed as limiting the present invention.

[0014] Example 1. Application of quercetin as an active ingredient in a cosmetic with skin oil-controlling effects.

[0015] The mass percentage of quercetin added to the cosmetic is 0.000025%-0.0004%.

[0016] Example 2. Application of quercetin as an active ingredient in a cosmetic with skin oil-controlling effects.

[0017] The quercetin is added to the cosmetic at a mass percentage of 0.0002%.

[0018] Verification of the efficacy of quercetin Experiment 1: Effect of quercetin on the activity of human sebaceous gland cells 1.1 Experimental Methods: A multi-functional microplate reader measures the absorbance at a wavelength of 490 nm. Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple formazan crystals, which are deposited in the cells. Dead cells do not have this function. Dimethyl sulfoxide (DMSO) solution can dissolve formazan in cells. Since the amount of crystal formation is directly proportional to the number of living cells, the absorbance value can indirectly reflect the number of living cells.

[0019] 1.2 Experimental Grouping 1.3 Experimental Procedure Human sebaceous gland cells were seeded in 96-well plates (5 × 10⁻⁶ cells / well). 3 (each well), incubated at 37°C for 24 h under 5% CO2 conditions; A blank solvent group and a blank control group were set up. Quercetin was dissolved in DMSO. Fresh culture medium containing 0.000025%, 0.00005%, 0.0001%, 0.0002%, 0.0004%, 0.0006%, and 0.0008% quercetin was added to the sample groups, respectively, and the samples were incubated at 37°C with 5% CO2 for 24 h. After incubation, add MTT solution to each well and continue culturing for 4 h; Remove the culture medium, add DMSO solution, shake to mix, and measure the absorbance at 490 nm. 1.4 Experimental Results The effects of quercetin on the activity of human sebaceous gland cells are shown in Table 2. The survival rate of human sebaceous gland cells treated with different concentrations of quercetin decreased in a dose-dependent manner with increasing drug concentration.

[0020] Compared with the control group, quercetin at concentrations below 0.0004% showed no significant difference in cell viability; however, when quercetin at concentrations of 0.0006% and 0.0008% was applied to human sebaceous gland cells, the cell viability was significantly reduced compared with the control group.

[0021] The results showed that quercetin had no cytotoxicity at concentrations below 0.0004%.

[0022] Experiment 2: Effects of quercetin on lipid accumulation in human sebaceous gland cells 2.1 Experimental Methods: Fluorescence intensity is detected by fluorescence microscopy. Nile Red is a lipophilic fluorescent dye that binds to neutral lipids (such as triglycerides) within cells and emits fluorescence. Its fluorescence intensity is directly proportional to the intracellular lipid content; therefore, intracellular lipid levels can be detected by fluorescence microscopy.

[0023] 2.2 Experimental Grouping 3. Experimental Procedure Human sebaceous gland cells were seeded in 12-well plates (1×10⁻⁶ cells / well). 5 (each well), incubated at 37°C for 24 h under 5% CO2 conditions; After incubation, remove the culture medium and wash with D-Hanks 1-2 times. The blank control group used fresh culture medium, the negative control group used fresh culture medium containing 0.0188 mM dihydrotestosterone, the sample group used culture medium containing dihydrotestosterone and the corresponding concentration of quercetin, and the positive control group used fresh culture medium containing dihydrotestosterone and 50 μg / mL tetrandrine. Incubation continued at 37℃, 5% CO2 for 24 h; each group had 3 replicates. After incubation, remove the culture medium, pre-cool, wash three times with D-Hanks, fix with 4% paraformaldehyde, wash again with D-Hanks, stain with Nile Red, take pictures with a fluorescence microscope, and analyze the average fluorescence intensity of each group using Image J. In the formula, A represents the average fluorescence intensity.

[0024] 2.4 Experimental Results The effects of quercetin on lipid accumulation in human sebaceous gland cells are shown in Table 4 and Appendix. Figure 2 As shown: Compared with the blank control group, ###p<0.001; compared with the negative control group, ***p<0.001. The fluorescence intensity of Nile Red binding to intracellular lipids showed that lipid accumulation significantly increased after treatment with the negative control group, while the addition of the positive control group resulted in a 40% inhibition rate of lipid accumulation, confirming the successful establishment of the model. Compared with the negative control group, treatment with 0.00005% quercetin inhibited lipid accumulation in sebaceous gland cells, with an inhibition rate of 12%; when the quercetin concentration was increased to 0.0002% and 0.0004%, lipid accumulation was significantly reduced, with an inhibition rate of approximately 40%. These results indicate that quercetin has a significant effect on inhibiting sebaceous gland secretion.

[0025] Experiment 3: Effects of quercetin on lipid synthesis and secretion regulators 3.1 Experimental Methods: The expression levels of lipid metabolism-related genes (acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase (SCD), insulin-like growth factor 1 receptor (IGF-1R), lipid droplet-coated protein 2 (PLIN2), and sterol regulatory element-binding protein 1C (SREBP-1C)) in human sebaceous gland cells were detected by RT-qPCR.

[0026] 3.2 Experimental Grouping 3.3 Experimental Procedure Human sebaceous gland cells were seeded in 6-well plates (3 × 10⁻⁶ cells / well). 5Incubate at 37℃ and 5% CO2 for 24 h; After incubation, the culture medium was removed, and the samples were washed 1-2 times with D-Hanks solution. The blank control group used fresh culture medium, the negative control group used fresh culture medium containing 0.0188 mM dihydrotestosterone, the sample group used culture medium containing dihydrotestosterone and 0.0002% quercetin, positive control group 1 used fresh culture medium containing dihydrotestosterone and 0.5 μg / mL claspone, and positive control group 2 used fresh culture medium containing dihydrotestosterone and 0.078 μM AKT inhibitor. Incubation was continued at 37°C and 5% CO2 for 24 h. Three replicates were set up for each group. After incubation, the culture medium was removed, the cells were pre-cooled, washed 1-2 times with D-Hanks, total RNA was extracted from each experimental group, cDNA was synthesized, and the gene expression of β-actin and the target gene was detected by q-PCR. Using β-actin as an internal reference for gene expression, the relative RNA expression level of the target gene was calculated.

[0027] In the formula, P represents the relative expression levels of the FAS, ACC, SCD, IGF-1R, PLIN2, and SREBP-1C genes.

[0028] 3.4 Experimental Results (1) Effect of quercetin on ACC gene expression: Compared with the blank control group, ##p<0.01; compared with the negative control group, **p<0.01 Acetyl-CoA carboxylase (ACC) is a biotin-dependent multifunctional enzyme and the rate-limiting step in fatty acid synthesis. It catalyzes the first step in fatty acid biosynthesis, namely the conversion of acetyl-CoA to malonyl-CoA. By inhibiting ACC to reduce fatty acid synthesis, ACC can also reduce lipid accumulation by regulating the AMPK / PPARα / CPT1A pathway; at the same time, inhibiting ACC can also enhance mitochondrial function and alleviate oxidative stress. Therefore, downregulating ACC expression can reduce lipid synthesis and achieve the goal of oil control; As shown in Table 6, the relative expression level of the ACC gene was significantly upregulated after stimulation with the negative control group, and the gene expression of ACC was downregulated by 40% after treatment with clavone, proving that the model was successfully constructed. Treatment with 0.0002% quercetin reduced the relative expression of the ACC gene by 34%, demonstrating that quercetin has a significant function in downregulating ACC gene expression and inhibiting lipid production at the gene regulation level.

[0029] (2) Effect of quercetin on FAS gene expression: Compared with the blank control group, ###p<0.001; compared with the negative control group, **p<0.01 Fatty acid synthase (FAS) is the main enzyme required for the synthesis of fatty acids from dietary carbohydrates. In the presence of NADPH, FAS catalyzes the conversion of acetyl-CoA and malonyl-CoA into long-chain saturated fatty acids, making it a key enzyme in fatty acid synthesis. Therefore, downregulating the high expression of the FAS gene can effectively reduce lipid synthesis; The relative expression levels of the FAS gene are shown in Table 7: After stimulation by the negative control group, FAS gene expression was significantly upregulated. After treatment with clavusone, the FAS gene expression was downregulated by 33% compared with the negative control group, proving that the model was successfully constructed. After treatment with 0.0002% quercetin, the relative expression level of the FAS gene was downregulated by 24%, demonstrating that quercetin has a strong ability to downregulate the expression of the FAS gene, showing its potential for oil control at the gene level.

[0030] (3) Effect of quercetin on SCD gene expression: Compared with the blank control group, ##p<0.001; compared with the negative control group, **p<0.01, ***p<0.001 Stearoyl-CoA desaturase (SCD) is a key enzyme that regulates lipid metabolism. It is responsible for catalyzing the conversion of saturated fatty acids into monounsaturated fatty acids and is the rate-limiting enzyme in the synthesis of monounsaturated fatty acids. The expression level of SCD directly affects the ratio of saturated fatty acids to monounsaturated fatty acids in the body, thereby further influencing lipid metabolism throughout the organism. During adipocyte differentiation, SCD also affects lipid droplet formation by regulating the expression of lipid synthesis-related genes. SCD can serve as a targeted therapy for drug intervention in lipid metabolism disorders. The RQ-PCR results (Table 8) showed that the negative control group significantly upregulated SCD gene expression, while the positive control group 1 was downregulated by 29% after treatment with clavone, proving that the model was successfully constructed. Treatment with 0.0002% quercetin had a more significant effect on downregulating the relative expression of the SCD gene, with a downregulation rate of 58%; this result indicates that quercetin can downregulate SCD expression at the gene level, thereby regulating lipid synthesis.

[0031] (4) Effect of quercetin on IGF-1R gene expression: Compared with the blank control group, #p<0.05; compared with the negative control group, *p<0.05, **p<0.01 Insulin-like growth factor 1 receptor (IGF-1R) is a transmembrane receptor that plays a role in cell proliferation, differentiation, and transformation. It is associated with the sebum secretion of sebaceous gland cells and can affect cell proliferation and differentiation, which are also related to the function of sebaceous gland cells. Activation of IGF-1R can promote cell proliferation and growth through specific signaling pathways such as MAPK and PI3K / AKT, which may indirectly increase sebum production by sebaceous gland cells. Therefore, downregulating the expression of IGF-1R can inhibit the excessive proliferation of sebaceous gland cells and achieve the purpose of oil control.

[0032] As shown in Table 9, the negative control group treatment led to a significant upregulation of IGF-1R gene expression, while the treatment with clavusone (positive control group 1) significantly downregulated IGF-1R expression by 27%, proving that the model was successfully constructed. Treatment with 0.0002% quercetin also significantly downregulated IGF-1R expression, with a downregulation rate of 30%; this result indicates that quercetin can downregulate IGF-1R expression and thus regulate sebum secretion.

[0033] (5) Effect of quercetin on PLIN2 gene expression: Compared with the blank control group, #p<0.05; compared with the negative control group, *p<0.05, **p<0.01 PLIN2 is a member of the lipid droplet coating protein (PLIN) family, also known as an adipocyte differentiation-associated protein, which participates in lipid metabolism and affects fat deposition. In sebaceous gland cells, PLIN2 may regulate sebum secretion by influencing lipid synthesis and catabolism. Therefore, downregulating PLIN2 expression can effectively reduce lipid droplet secretion. The relative expression levels of the PLIN2 gene are shown in Table 10. Under the stimulation of the negative control group, the expression of PLIN2 gene was significantly upregulated. Treatment with clavone (positive control group 1) can significantly reduce the stimulation and downregulate the expression of PLIN2 gene by 37%, proving that the model was successfully constructed. Treatment with 0.0002% quercetin significantly downregulated PLIN2 expression by 30%, demonstrating that quercetin has the effect of downregulating PLIN2 expression and thus inhibiting sebaceous gland secretion.

[0034] (6) Effect of quercetin on SREBP-1C gene expression: Compared with the blank control group, #p<0.05; compared with the negative control group, *p<0.05, ***p<0.001 SREBP-1C belongs to the sterol regulatory element-binding protein (SREBP) family. The lipid metabolism pathway mediated by this family is the primary link in the study of lipid metabolism regulation mechanisms, responsible for regulating processes such as exogenous lipid uptake and internal lipid synthesis. SREBP-1C is a key regulator of lipid metabolism signal transduction, and its target genes mainly include FAS, SCD, acetyl-CoA synthase (ACS), and hydroxymethylglutaryl-CoA reductase (HMGCR). Therefore, reduced SREBP-1C activity can inhibit key enzymes in lipid synthesis, thereby controlling the downregulation of sebum secretion.

[0035] Table 11 shows the RQ-PCR results of SREBP-1C. Stimulation of the negative control group led to a significant upregulation of SREBP-1C gene expression, while treatment with the AKT inhibitor (positive control group 2) downregulated SREBP-1C gene expression by 19%, proving that the model was successfully constructed. Treatment with 0.0002% quercetin significantly downregulated the relative expression of the SREBP-1C gene, with a downregulation rate of 63%, demonstrating that quercetin can inhibit lipid synthesis at the gene level and achieve oil control.

[0036] In summary, based on human sebaceous gland cells, quercetin at a concentration of 0.0002% significantly downregulated ACC, FAS, SCD, IGF-1R, PLIN2, and SREBP-1C, especially SCD, SREBP-1C, and PLIN2. Therefore, it can be demonstrated that quercetin can better regulate the expression of genes related to lipid synthesis and has the effect of inhibiting sebaceous gland secretion and controlling oil production in the skin.

Claims

1. The application of quercetin as an active ingredient in cosmetics with skin oil-controlling effects.

2. The application of quercetin according to claim 1, characterized in that: The mass percentage of quercetin added to the cosmetic is 0.000025%-0.0004%.

3. The application of quercetin according to claim 2, characterized in that: The quercetin is added to the cosmetic at a mass percentage of 0.0002%.

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