Application of glycerol glucoside in the preparation of oil control products

Abstract

The present invention relates to an application of glycerol glucoside in the preparation of oil control products, belonging to the technical field of cosmetics. The glycerol glucoside of the present invention can reduce PPAR-γ Genes and / or MC‑1R The expression of the gene can be reduced, thereby reducing the synthesis of lipid droplets in sebaceous gland cells, fundamentally inhibiting oil secretion and achieving an oil control effect. It can be widely used in the preparation of oil-control skin care products, cosmetics, and cleaning products, or in the preparation of products for the treatment or improvement of PPAR-γ Genes and / or MC‑1R Drugs for diseases related to gene expression.

Description

Technical Field

[0001] The invention relates to application of glycerol glucoside in preparing an oil-control product, and belongs to the technical field of cosmetics. Background Art

[0002] Human sebaceous glands (SG) are microscopic, branching multi-glandular organs that are important appendages of the skin and are located in the dermis and open to the skin surface. They are distributed on the surface of the skin and their main function is to produce and secrete an oily, waxy substance (sebum) by differentiating and dividing fully mature cells, a unique process called holocrine secretion. Sebaceous glands are rich in capillaries and nerve endings that respond to external stimuli and affect the synthesis and secretion of skin lipids. In individuals with oily skin, the secretion of sebaceous glands in the T zone increases, leading to skin diseases such as acne, rosacea, and seborrheic dermatitis.

[0003] The main factors that affect sebum secretion include age, gender, endocrine (androgen), growth factors, diet and season. Skin oil control can be achieved through oral and topical products. There are three modes of action for oil control through topical medications or cosmetics: (1) temporarily removing or absorbing excess sebum; (2) inhibiting the spread of sebum; (3) reducing sebum synthesis. Removing or absorbing excess sebum and inhibiting the spread of sebum can only temporarily solve the effects caused by excessive sebum secretion, and it is not a cure for the cause. Only by inhibiting the function of sebaceous glands can the effect of reducing oil secretion lastingly and completely be achieved.

[0004] The lipid droplet synthesis process includes the following four steps: (1) synthesis of triglycerides in the endoplasmic reticulum; (2) formation of oil vesicles in the endoplasmic reticulum membrane; (3) budding and formation of new lipid droplets; (4) growth and expansion of lipid droplets through specific proteins. Lipid droplets are regulated by a large number of proteins. The most relevant membrane protein is the perilipin (PLIN) family, which has structural and regulatory properties. PLIN1 and PLIN2 are expressed in hamster sebaceous gland cells, and PLIN3 has been shown to regulate specific lipogenesis pathways in human oil cells. Downregulation of PLIN3 disrupts lipid accumulation and changes the expression of key enzymes for sebaceous gland lipogenesis, indicating that PLIN3 affects pathways related to sebaceous gland lipogenesis, such as desaturation and triglyceride synthesis. In addition, human sebaceous gland cells express nuclear retinoic acid receptors RAR γ, RAR α and RXR α. Each sebaceous gland cell has about 1500 nuclear receptor molecules, which regulate the amount of sebum secreted by sebaceous gland cells.

[0005] PPAR is a member of the nuclear hormone receptor (NHR) family and acts as a transcriptional regulator of multiple genes, including genes involved in lipid metabolism in adipose tissue, liver, and skin, by binding to RXR α. PPAR is expressed in human SGs and human SZ95 sebocytes.PPAR-γ It can induce differentiation and lipid production of human sebocytes. MC-1R ) is an important membrane protein in sebum differentiation and synthesis. Melanocortin initiates intracellular signaling by binding to and activating g-protein-coupled melanocortin receptors (MCRs), thereby increasing the production of cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) activity. cAMP and PKA promote sebum synthesis by regulating the activity and gene expression of lipid synthesis-related enzymes. For oil-control actives, the most typical one is isotretinoin, which is the most effective lipid synthesis inhibitor and can reduce the synthesis of triglycerides, wax / sterol esters, free fatty acids and squalene. Isotretinoin can bind to nuclear receptor molecules on sebaceous gland cells, thereby regulating the synthesis and metabolism of sebum and reducing the content of sebum. However, the use of isotretinoin often leads to adverse reactions such as dryness of the skin and mucous membranes, peeling, and skin sensitivity.

[0006] Current studies have shown that glycerol glucoside has unique moisturizing properties, can regulate skin hydration, and has a positive effect on the physiological functions of the skin; in addition, it has good biocompatibility, stability, anti-inflammatory and soothing properties, and other potential health care and therapeutic effects. Chinese patent document CN104651287A (application number 201310598111.2) discloses an engineered bacterium that can use solar energy to fix carbon dioxide under light, and can synthesize, accumulate and secrete glycerol glucoside under salt stress, increasing the yield to about 4 times that of the wild type. Therefore, with the increase in the yield of glycerol glucoside, combined with its many advantages, it is of great significance to explore the application of glycerol glucoside in other aspects. Summary of the invention

[0007] In view of the deficiencies in the prior art, the present invention provides an application of glycerol glucoside in the preparation of an oil control product.

[0008] The technical solution of the present invention is as follows: Application of glycerol glucoside or a composition containing glycerol glucoside in the preparation of an oil control product.

[0009] Preferably according to the present invention, the stereo configuration of the glycerol glucoside is 2-α GG.

[0010] According to the preferred embodiment of the present invention, the glycerol glucoside can downregulate PPAR-γ Genes and / or MC-1R The expression of genes can be reduced, thereby reducing the synthesis of lipid droplets in sebaceous gland cells to achieve the effect of oil control.

[0011] A kind of glycerol glucoside or the composition containing glycerol glucoside is used for preparing down-regulation PPAR-γ Genes and / or MC-1R Application of gene expression products.

[0012] A glycerol glucoside or a composition containing glycerol glucoside is used in the preparation of a composition for treating or improving PPAR-γ Genes and / or MC-1R Application of drugs in diseases related to gene expression.

[0013] According to the present invention, the treatment or improvement is preferably achieved by downregulating PPAR-γ Genes and / or MC-1R Gene expression is achieved.

[0014] An oil control product comprising an effective amount of glyceryl glucoside.

[0015] Preferably according to the present invention, the oil control product includes skin care products, cosmetics, cleaning products or medicines.

[0016] Further preferably, the oil control product can be applied externally to the skin of the face or head.

[0017] According to the preferred embodiment of the present invention, the oil control product can reduce PPAR-γ Genes and / or MC-1R The expression of genes can be reduced, thereby reducing the synthesis of lipid droplets in sebaceous gland cells to achieve the effect of oil control.

[0018] Beneficial effects: The glycerol glucoside of the present invention can downregulate PPAR-γ Genes and / or MC-1R The expression of the gene can be reduced, thereby reducing the synthesis of lipid droplets in sebaceous gland cells, fundamentally inhibiting oil secretion and achieving an oil control effect. It can be widely used in the preparation of oil-control skin care products, cosmetics, and cleaning products, or in the preparation of products for the treatment or improvement of PPAR-γ Genes and / or MC-1R Drugs for diseases related to gene expression. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Cell viability curves of samples with different concentrations; Figure 2 This is the result of Oil Red O staining; Figure 3 This is a bar graph of Oil Red O staining results; Figure 4 for PPAR-γ Gene expression bar graph; Figure 5 for MC-1R Gene expression bar graph. DETAILED DESCRIPTION

[0020] The technical solution of the present invention is further described below in conjunction with embodiments, but the scope of protection of the present invention is not limited thereto.

[0021] The contents not described in detail in the examples are all based on the existing conventional techniques in the art, and the experimental materials and reagents not described in detail are all common commercial products. Among them, the cells used in the examples are sebaceous gland cells, provided by Guangdong Boxi Biotechnology Co., Ltd., batch number: 210829. MEM culture medium was purchased from Gibco, PBS buffer (pH 7.2-7.4) was purchased from Solebao, DMSO and MTT were purchased from Sigma, fetal bovine serum (FBS) was purchased from Lanzhou Rongye, isotretinoin was purchased from Aladdin, Oil Red O was purchased from Solebao, hematoxylin staining solution was purchased from Biyuntian, paraformaldehyde was purchased from Biosharp, RNAiso Plus was purchased from Aikerui Biological, and reverse transcription kits and fluorescent dyes were purchased from Aikerui Biological.

[0022] In the following examples, the name of the experimental sample is Annuoyin, and the ingredients are: 50% glycerol glucoside, 45% water, and 5% pentanediol (preservative). Among them, the stereo configuration of glycerol glucoside is 2-α GG, and it is purchased from the blue algae extracted glycerol glucoside product produced by Qingdao Zhongke Lanzhi Biotechnology Development Co., Ltd., and the trade name is Annuoyin ® .

[0023] Example 1 Cytotoxicity test (1) Cell inoculation: Inoculate cells into a T75 flask containing cell culture medium and place in a 37°C, 5% CO2 incubator until the cells recover. Continue culturing until the plating rate reaches about 60%, and then inoculate at 1.1×10 4 The cells were seeded into a 96-well plate at a density of 100 cells / well and incubated overnight in a 37°C, 5% CO2 incubator.

[0024] (2) Experimental grouping: The experiment set up a zero adjustment group, a solvent control group, a positive control group and a sample group. In the sample group, each sample was set with 8 concentration gradients, and 3 replicate wells were set under each concentration gradient.

[0025] (3) Solution preparation: Prepare sample working solutions of different concentrations according to Table 1, using cell culture medium as the solvent.

[0026] Table 1. Test concentration setting table

[0027] (4) Dosing: Dosing was performed when the cell plating rate in the 96-well plate reached 40%-60%. In the solvent control group, 200 μL of cell culture medium was added to each well; in the positive control group, 200 μL of cell culture medium containing 10% DMSO was added to each well; in the sample group, 200 μL of cell culture medium containing samples of corresponding concentrations was added to each well; in the zero adjustment group, no cells were inoculated, and only 200 μL of cell culture medium was added. After dosing, the 96-well plate was placed in a 37°C, 5% CO2 incubator for 24 hours.

[0028] The above cell culture medium is MEM culture medium.

[0029] (5) Detection: After incubating the cells for 24 hours, discard the supernatant, add MTT working solution (0.5 mg / mL), and incubate at 37°C in the dark for 4 hours. After the incubation, discard the supernatant, add 150 μL DMSO to each well, and read the OD value at 490 nm. Calculate the relative cell viability according to the formula.

[0030] Relative cell viability (%) =

[0031] (6) The test results are shown in Table 2. The cell viability of samples with different concentrations is shown in Figure 1 As shown, the results show that Anuoyin has little cytotoxicity at low concentrations. In the following examples, a safe concentration of 2% was selected for experiments.

[0032] Table 2. Cell viability test results

[0033] Example 2 Oil content test (1) Cell inoculation: Inoculate cells into a T75 flask containing cell culture medium and place in a 37°C, 5% CO2 incubator until the cells recover. Continue culturing until the plating rate reaches about 60%, and then add 7.0×10 4 The cells were inoculated into 24-well plates at a density of 1000 cells / well and incubated overnight in a 37°C, 5% CO2 incubator.

[0034] (2) Solution preparation: Prepare the working solution of the test substance according to the test groups in Table 3, using cell culture medium as the solvent.

[0035] Table 3. Test groups

[0036] (3) Drug administration: When the cell plating rate in the 24-well plate reaches 40%~60%, group drug administration is carried out according to Table 3, with 3 replicate wells in each group. 1 mL of cell culture medium is added to each well of the blank control group, 1 mL of cell culture medium containing isotretinoin is added to each well of the positive control group, and 1 mL of cell culture medium containing 2% anorin is added to each well of the sample group. After drug administration, the 24-well plate is placed in a 37°C, 5% CO2 incubator for 72 hours, and the medium is changed and drug administration is performed every day during the culture period.

[0037] (4) Oil Red O staining: Aspirate and discard the old solution, wash with PBS buffer, fix the cells with paraformaldehyde, add 300 μL of Oil Red O working solution to each well, stain for 15 min in the dark, wash with PBS buffer, observe under a microscope, and calculate the integrated optical density (IOD).

[0038] (5) Calculate the lipid inhibition rate and compare the groups using t -test Statistical analysis, all tests are two-tailed. P <0.05 was considered to be significantly different. P <0.01 was considered to be extremely significant.

[0039] Inhibition rate (%) =

[0040] Note: Integrated optical density (IOD), its value reflects the oil content.

[0041] (6) Oil Red O staining results are as follows Figure 2 The results of Oil Red O staining are shown in Table 4, and the relative IOD average values ​​are shown in Table 4. Figure 3 As shown, the results showed that compared with the BC group, the oil content in the PC group decreased significantly, indicating that the positive control of this test was effective. Compared with the BC group, the oil content of the sample group with 2% concentration of Annoin decreased significantly, and the inhibition rate was 31.00%.

[0042] Table 4. Summary of Oil Red O staining results

[0043] Note: Use t When the -test method is used for statistical analysis, the significance compared with the BC group is indicated by *. P -value<0.05 is represented by *, P -value<0.01 is indicated as **.

[0044] Example 3 Gene expression test (1) Cell inoculation: Inoculate cells into a T75 flask containing cell culture medium and place in a 37°C, 5% CO2 incubator until the cells recover. Continue culturing until the plating rate reaches about 60%, and then inoculate at 3.0×10 5 The cells were inoculated into 6-well plates at a density of 1000 cells / well and incubated overnight in a 37°C, 5% CO2 incubator.

[0045] (2) Solution preparation: Prepare the working solution of the test substance according to the test groups in Table 5, using cell culture medium as the solvent.

[0046] Table 5. Test groups

[0047] (3) Drug administration: When the cell plating rate in the 6-well plate reaches 40%-60%, the drugs are administered in groups according to Table 5. Each group has 3 replicate wells. 2 mL of cell culture medium is added to each well of the blank control group, 2 mL of cell culture medium containing isotretinoin is added to each well of the positive control group, and 2 mL of cell culture medium containing 2% anorin is added to each well of the sample group. After the drug administration is completed, the 6-well plate is placed in a 37°C, 5% CO2 incubator for 72 hours. During the culture period, the medium is changed and the drug administration is performed every day.

[0048] (4) Collect cells: After incubation, discard the old solution, wash twice with PBS buffer, add 1 mL of RNAisoPlus to each well, pipette and lyse the cells, and then collect the samples.

[0049] (5) Gene expression detection: RNA was extracted, reverse transcribed into cDNA, and then fluorescent quantitative PCR was performed. Methods The results were calculated using GraphPad Prism and the results were expressed as Mean ± SD. t -test Statistical analysis. All statistical analyses were two-tailed. P <0.05 was considered to be significantly different. P The difference was considered to be extremely significant when the value was less than 0.01. The gene expression inhibition rate was calculated.

[0050] Inhibition rate (%) =

[0051] (6) PPAR-γ The test results are shown in Table 6. The gene expression levels are shown in Figure 4 As shown, MC-1R The test results are shown in Table 7. The gene expression levels are shown in Figure 5 The results show that PPAR-γ、MC-1R The gene expression levels were significantly downregulated, with inhibition rates of 28.00% and 28.00% respectively, indicating that the sample at a concentration of 2% can reduce the synthesis of lipid droplets in sebaceous gland cells and downregulate PPAR-γ、MC-1R Gene expression level, has oil control effect.

[0052] Table 6. PPAR-γ Summary table of genetic test results

[0053] Table 7. MC-1R Summary table of genetic test results

[0054] Note: Use The method is used to calculate the results. t When the -test method was used for statistical analysis, the mRNA amplification fold of the BC group was normalized, and the significance was indicated by * compared with the BC group. P -value<0.05 is represented by *, P -value<0.01 is indicated as **.

Claims

1. Use of glycerol glucoside or a composition comprising glycerol glucoside in the preparation of an oil control product.

2. The use according to claim 1, characterized in that The stereo configuration of the glycerol glucoside is 2-α GG.

3. The use according to claim 1, characterized in that The glycerol glucoside can downregulate PPAR-γ Genes and / or MC-1R The expression of genes can be reduced, thereby reducing the synthesis of lipid droplets in sebaceous gland cells to achieve the effect of oil control.

4. A method for preparing a glycerol glucoside or a composition comprising glycerol glucoside PPAR-γ Genes and / or MC-1R Application of gene expression products.

5. A glycerol glucoside or a composition comprising glycerol glucoside for use in the treatment or improvement of PPAR-γ Genes and / or MC-1R Application of drugs in diseases related to gene expression.

6. The use according to claim 5, characterized in that The treatment or improvement is achieved by downregulating PPAR-γ Genes and / or MC-1R Gene expression is achieved.

7. An oil control product, characterized in that: Includes an effective amount of glyceryl glucoside.

8. The product according to claim 7, characterized in that Including skin care products, cosmetics, cleaning products or medicines.

9. The product according to claim 8, characterized in that Can be applied externally to the face or scalp.

10. The product according to claim 7, characterized in that Can be adjusted down PPAR-γ Genes and / or MC-1R The expression of genes can be reduced, thereby reducing the synthesis of lipid droplets in sebaceous gland cells to achieve the effect of oil control.

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