Stimulation-reducing and anti-aging retinoid composition and application thereof

Abstract

The invention discloses a retinoid composition capable of reducing irritation and resisting aging and application thereof, and belongs to the technical field of preparations for dressing and makeup. The irritation-reducing and anti-aging retinoid composition comprises an anti-aging component and ginsenoside CK, the anti-aging component comprises at least one of retinol and a retinol derivative, and the mass ratio of the anti-aging component to the ginsenoside CK is 1: (0.001-2). The retinoid composition disclosed by the invention has the effects of being small in irritation and synergistically promoting aging resistance. According to the invention, the verbascos exosome is further compounded with a retinoid composition, and the retinoid composition enters the exosome through ultrasonic treatment and incubation so as to enable vesicles to wrap the composition, so that the activity of the composition can be protected, the irritation is further reduced, and the anti-aging effect is improved. Therefore, the retinoid composition provided by the invention is suitable for daily cosmetics.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetic preparations, and in particular relates to a retinoid composition for reducing irritation and resisting aging and an application thereof. Background Art

[0002] Skin aging is a complex process, which can be divided into endogenous aging and exogenous aging. Endogenous factors include gender and genetic variation. Exogenous factors include ultraviolet radiation and air pollution, some chronic diseases and lifestyle choices. Although aging is inevitable, premature aging can be avoided by reducing or resisting external stimuli. Exogenous skin aging caused by long-term ultraviolet radiation is called photoaging. Due to the loss of collagen, the destruction of elastic fibers and collagen fibers, the skin will show symptoms of wrinkles, sagging and pigmentation, and in severe cases, it will cause inflammation and skin cancer. The ultraviolet rays that reach the earth's surface from the sun are mainly composed of long-wave ultraviolet rays (UVA) (wavelength 320-400nm) and medium-wave ultraviolet rays (UVB) (wavelength 280-320nm). Compared with UVB, UVA has stronger penetration and can reach the dermis, destroy collagen fibers, cause skin aging, and produce wrinkles.

[0003] The external manifestations of skin aging include skin sagging, drying, thinning, loss of luster, wrinkle formation, and pigmentation. The internal manifestations include thinning of the epidermis, decreased hydration, dry skin, and loss of collagen and elastin in the dermis that maintain skin elasticity and firmness. Therefore, the efficacy of anti-aging cosmetics mainly revolves around enhancing the skin's proliferation and metabolism capacity, rebuilding the skin's extracellular matrix, resisting ultraviolet radiation, anti-oxidation, and anti-allergic inflammation.

[0004] Retinol is a commonly used anti-aging substance for the skin. It can reduce the formation of wrinkles, increase the thickness of the epidermis and dermis, and delay skin aging by topical application or oral administration. However, when retinol is used alone, skin reactions such as redness, scaling, and inflammation will occur. In addition, retinol is unstable and easily oxidized by light, which is very irritating. Compared with retinol, the irritation of retinol derivatives (such as hydroxypinacolone retinoic acid ester (HRP) and retinyl palmitate (VAPA)) is lower, but its activity is relatively reduced, and the anti-aging effect is reduced. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a retinoid composition that can reduce irritation and has excellent anti-aging effects.

[0006] The present invention provides a retinoid composition for reducing irritation and anti-aging, comprising an anti-aging component and ginsenoside CK, wherein the anti-aging component comprises at least one of retinol and a retinol derivative, and the mass ratio of the anti-aging component to the ginsenoside CK is 1:0.001-2.

[0007] As a preferred embodiment of the present invention, when the anti-aging ingredient is retinol, it is further preferred that the mass ratio of retinol to ginsenoside CK in the retinoid composition is 1:0.02 to 2. More preferably, the mass ratio of retinol to ginsenoside CK in the retinoid composition is 1:0.1 to 1.

[0008] As a preferred embodiment of the present invention, the retinol derivative includes at least one of hydroxypinacolone retinoic acid ester, retinol palmitate, retinol propionate, retinol retinoic acid ester, retinol linoleate, and retinol acetate.

[0009] Furthermore, when the anti-aging ingredient is hydroxypinacolone retinoic acid ester, the mass ratio of hydroxypinacolone retinoic acid ester to ginsenoside CK in the retinoid composition is preferably 1:0.002-0.5, and more preferably the mass ratio of hydroxypinacolone retinoic acid ester to ginsenoside CK in the retinoid composition is 1:0.02-0.2.

[0010] Furthermore, when the anti-aging ingredient is retinol palmitate, the mass ratio of retinol palmitate to ginsenoside CK in the retinoid composition is preferably 1:0.002-0.5, and the mass ratio of retinol palmitate to ginsenoside CK in the retinoid composition is more preferably 1:0.01-0.1.

[0011] The preparation method of the retinoid composition is as follows: uniformly mixing the components of the retinoid composition to obtain the retinoid composition.

[0012] The present invention also provides application of the retinoid composition in preparing daily cosmetics.

[0013] As a preferred embodiment of the present invention, the mass percentage of the retinoid composition in the daily cosmetics is 0.01% to 10%.

[0014] The inventor unexpectedly discovered during the research process that by adding the composition solution dropwise and then performing ultrasonic treatment and incubation, the retinoid composition can enter the vesicle structure of the verbascosa exosomes, achieving the encapsulation of the retinoid composition by the natural vesicle structure, which can not only effectively reduce irritation, but also significantly enhance the anti-aging effect. Therefore, the present invention further provides a retinoid composition preparation, which includes the above-mentioned retinoid composition and verbascosa exosomes, and the mass ratio of the retinoid composition to the verbascosa exosomes is 1 to 120:1.

[0015] As a preferred embodiment of the present invention, in the above-mentioned retinoid composition preparation, the mass ratio of the retinoid composition to the verbabasin exosomes is preferably 2 to 15:1.

[0016] The above-mentioned mullein exosomes are prepared by the following method: mullein and PBS buffer are mixed and then ground, filtered with gauze, and the filtrate is collected; the collected filtrate is centrifuged at a centrifugal force of 1000-3000g for 20-60min, and the supernatant is taken; the supernatant is centrifuged at a centrifugal force of 100000-200000g for 60-120min, the supernatant is discarded, the precipitate is taken, and the mullein exosomes are obtained by freeze-drying.

[0017] The preparation method of the above-mentioned retinoid composition preparation is: dispersing the retinoid composition in butanediol to obtain a composition solution; after mixing the verbascosum exosomes and PBS buffer, dripping the composition solution, ultrasonically treating, incubating, centrifuging, and freeze-drying, to obtain the retinoid composition preparation.

[0018] As a preferred embodiment of the present invention, in the method for preparing the retinoid composition preparation, the droplet acceleration rate of the composition solution is 1 to 10 mL / min.

[0019] As a preferred embodiment of the present invention, in the method for preparing the above-mentioned retinoid composition preparation, the power of the ultrasonic treatment is 200 to 500 W, and the ultrasonic time is 1 to 5 min.

[0020] As a preferred embodiment of the present invention, in the method for preparing the above-mentioned retinoid composition preparation, the incubation temperature is room temperature, and the incubation time is 12 to 24 hours.

[0021] The present invention also provides the use of the above retinoid composition preparation in the preparation of daily cosmetics.

[0022] As a preferred embodiment of the present invention, the daily cosmetics include any one of facial cleanser, shower gel, skin care lotion, skin care cream, essence, eye cream, and facial mask.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] 1. The retinoid composition of the present invention compounds retinol or / and retinol derivatives with ginsenoside CK, which significantly reduces the irritation of retinol and retinol derivatives, has low cytotoxicity, and improves cell survival rate. In daily cosmetics, the irritation of retinol and its derivatives can be reduced by compounding with ginsenoside CK, thereby reducing skin problems such as desquamation and redness caused by their use in daily cosmetics.

[0025] 2. The retinoid composition of the present invention compounds retinol or / and retinol derivatives with ginsenoside CK, which has a significant proliferation-promoting effect on photoaged HaCaT cells, and the two have a coordinated synergistic effect, thereby improving the anti-aging effect, and can alleviate the senescence of photoaged HaCaT cells induced by UVA radiation, significantly reduce UVA-induced HaCaT cell apoptosis, promote the expression of collagen and elastin, and inhibit the oxidative stress response of cells; therefore, the retinoid composition provided by the present invention is suitable for daily cosmetics.

[0026] 3. The present invention uses mullein exosomes and a retinoid composition to compound. The retinoid composition enters the exosomes through ultrasound and incubation so that the vesicles wrap the composition, which can protect the activity of the composition, further reduce irritation, and improve the anti-aging effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a bar graph of the cell survival rate of the combination of VA and ginsenoside CK in Example 1.

[0028] Figure 2 It is a bar graph of the cell survival rate of the complex of HPR and ginsenoside CK in Example 1.

[0029] Figure 3 It is a bar graph of the cell survival rate of the combination of VAPA and ginsenoside CK in Example 1.

[0030] Figure 4 It is a bar graph of cell survival rates under different radiation doses in Example 2.

[0031] Figure 5 This is a photo of β-galactosidase staining in Example 2.

[0032] Figure 6 This is a photograph showing the development of photoaging cell apoptosis under the combined drug use in Example 2.

[0033] Figure 7 It is a bar graph of the expression levels of apoptosis-related proteins under the combined drug use in Example 2.

[0034] Figure 8 This is a photograph of the development of collagen and elastin in photoaged cells under the combined use of drugs in Example 2.

[0035] Fig. 9 It is a bar graph of the expression levels of collagen and elastin under the combined drug use in Example 2. DETAILED DESCRIPTION

[0036] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments and comparative examples, the purpose of which is to understand the content of the present invention in detail, rather than to limit the present invention. All other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0037] Unless otherwise specified, the experimental reagents and instruments involved in the embodiments of the present invention are all commonly used common reagents and instruments. The pH value of the following PBS buffer is 7.0.

[0038] Example 1

[0039] 1. The irritation-reducing effect of retinol or / and retinol derivatives combined with ginsenoside CK

[0040] The MTT method was used to test the effect of drugs on the proliferation of HaCaT cells. The test drugs were: retinol (VA), hydroxypinacolone retinoic acid ester (HPR), retinol palmitate (VAPA) and ginsenoside CK. The MTT method includes the following steps:

[0041] HaCaT cells in the logarithmic growth phase were digested with trypsin and centrifuged; the cells were centrifuged at a rate of 1.5×10 4 The cells were plated at a density of 100 cells / well in a 96-well plate, 0.1 mL / well of cell suspension was added, and the culture plate was placed in an incubator (37°C, 5% CO 2 ) for 24 hours; after the cells adhered to the wall, the medium in the 96-well plate was discarded, and different concentrations of drugs were added for 48 hours, and then 20 μL / well of MTT solution (5 mg / mL) was added, and incubated for 3-4 hours. 150 μL of blue-purple formazan dissolved in DMSO was added to each well; the wavelength of 490 nm was selected, and the light absorption value of each well was measured on an enzyme-linked immunosorbent assay, and the results were recorded. The cell survival rate was calculated according to the formula: Inhibition rate (%) = [(OD 阴性对照孔 -OD 实验孔 ) / (OD 阴性对照孔 -OD 空白孔 )]×100%, survival rate=100%-inhibition rate, and graphing was performed using GraphPad Prism9 software. The results are shown in Figures 1 to 3 shown.

[0042] The experimental results showed that when VA was administered alone at a concentration of 0.025 mg / mL and 0.05 mg / mL, the cell survival rates were 51.99% and 44.26%, respectively. When it was compounded with different concentrations of ginsenoside CK (0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL and 0.05 mg / mL), the cell survival rates increased to 94.24% to 109.77%. When HPR was administered alone at a concentration of 0.1 mg / mL, 0.25 mg / mL and 0.5 mg / mL, the cell survival rates were 62.70%, 14.38% and 16.02%, respectively. When it was compounded with different concentrations of ginsenoside CK (0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL and 0.05 mg / mL), the cell survival rates were significantly increased. When VAPA was administered alone at concentrations of 0.1, 0.25 mg / mL and 0.5 mg / mL, the cell survival rates were 64.05%, 76.96% and 78.70%, respectively. When it was compounded with different concentrations of ginsenoside CK (0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL and 0.05 mg / mL), the cell survival rates were significantly improved. This indicates that compounding retinol or / and retinol derivatives with ginsenoside CK significantly reduces the irritation of retinol and retinol derivatives.

[0043] 2. Anti-aging properties of retinol or / and retinol derivatives combined with ginsenoside CK

[0044] (1) Establishing a photoaging model

[0045] The photoaging model is constructed using UVA with a wavelength of 365 nm. The construction method includes the following steps:

[0046] HaCaT cells in the logarithmic growth phase were digested with trypsin and centrifuged; the cells were centrifuged at a rate of 1.5×10 4 The cells were plated at a density of 100 cells / well in a 96-well plate, 0.1 mL / well of cell suspension was added, and the culture plate was placed in an incubator (37°C, 5% CO 2 ) for 24 h. After the cells adhered, the medium in the 96-well plate was discarded, and the cells were washed twice with PBS buffer and the washing solution was discarded. Then, an appropriate amount of PBS buffer was added to cover the cells to prevent the cells from drying out. The UVA irradiation power was 10 mW / cm 2 The irradiation distance was 10 cm, and the UVA radiation doses of each group were 0 and 10 J / cm 2 , 20J / cm 2 、30J / cm 2 , 40J / cm 2 , 50J / cm 2 , according to the formula: Radiation dose (J / cm 2) = UVB irradiation power (W / cm 2 )×time (s) to calculate the corresponding irradiation time. After irradiation, remove PBS, add 20 μL / well of MTT solution (5 mg / mL), incubate for 3-4 hours, add 150 μL of blue-purple formazan dissolved in DMSO to each well; select 490 nm wavelength, measure the light absorption value of each well on the enzyme-linked immunosorbent monitor, and record the results.

[0047] According to the formula: Inhibition rate (%) = [(OD 阴性对照孔 -OD 实验孔 ) / (OD 阴性对照孔 -OD 空白孔 )] × 100%, survival rate = 100% - inhibition rate, calculate cell survival rate, and use GraphPad Prism9 software to process and draw graphs. The results are shown in Figure 4 shown.

[0048] from Figure 4 It can be seen that with the increase of UVA irradiation dose, the viability of HaCaT cells weakened. The radiation dose that inhibited the viability of HaCaT cells by 50% was selected for subsequent experiments, i.e., 40 J / cm 2 Radiation dose.

[0049] (2) Test drug cytotoxicity

[0050] The MTT method was used to test the effect of drugs on the proliferation of HaCaT cells. The test drugs were: retinol (VA), hydroxypinacolone retinoic acid ester (HPR), retinol palmitate (VAPA) and ginsenoside CK. The MTT method includes the following steps:

[0051] HaCaT cells in the logarithmic growth phase were digested with trypsin and centrifuged; the cells were centrifuged at a rate of 1.5×10 4 The cells were plated at a density of 100 cells / well in a 96-well plate, 0.1 mL / well of cell suspension was added, and the culture plate was placed in an incubator (37°C, 5% CO 2 ) for 24 h; after the cells adhered, the medium in the 96-well plate was discarded, and the cells were washed twice with PBS buffer and the washing solution was discarded. Then, an appropriate amount of PBS buffer was added to cover the cells to prevent the cells from drying out. The UVA irradiation power was 10 mW / cm 2 , the irradiation distance is 10cm, and the UVA radiation dose is 40J / cm 2, the irradiation time is 66.67min. Then, the drugs are administered according to the grouping and the drug administration concentration of Tables 1 to 4. After the drug acts for 24h, 20μL / well of MTT solution (5mg / mL) is added, incubated for 3 to 4h, and 150μL of DMSO-dissolved blue-purple formazan is added to each well; a wavelength of 490nm is selected, and the light absorption value of each well is measured on an enzyme-linked immunosorbent monitor, and the results are recorded. The cell survival rate is calculated according to the formula of the embodiment. Among them, A1, B1, C1 and D1 are blank control groups, and the blank control groups are not subjected to UVA irradiation and no drug administration; A2, B2, C2 and D2 are irradiation control groups, and the irradiation control groups are only subjected to UVA irradiation without drug administration; A3 to A6, B3 to B9, C3 to C8 and D3 to D7 are drug administration groups, and the drug administration groups are subjected to UVA irradiation and drug administration at the same time. The experimental results are shown in Tables 1 to 4.

[0052] Table 1 Effect of ginsenoside CK alone on the proliferation of UVA-irradiated HaCaT cells

[0053]

[0054] Table 2 Effect of combined administration of VA and ginsenoside CK on proliferation of HaCaT cells irradiated by UVA

[0055]

[0056] Table 3 Effect of combined administration of HPR and ginsenoside CK on proliferation of HaCaT cells irradiated by UVA

[0057]

[0058] Table 4 Effect of combined administration of VAPA and ginsenoside CK on proliferation of HaCaT cells irradiated by UVA

[0059]

[0060]

[0061] (3) The β-galactosidase staining method was used to detect the effect of the combined administration of anti-aging ingredients (retinol, HPR and VAPA) and ginsenoside CK on the senescence level of photoaged cells; the β-galactosidase staining method included the following steps:

[0062] HaCaT cells in the logarithmic growth phase were digested with trypsin and centrifuged; the cells were centrifuged at a rate of 1.5×10 4 The cells were plated at a density of 100 cells / well in a 96-well plate, 0.1 mL / well of cell suspension was added, and the culture plate was placed in an incubator (37°C, 5% CO 2) for 24 h; after the cells adhered, the medium in the 96-well plate was discarded, and the cells were washed twice with PBS buffer and the washing solution was discarded. Then, an appropriate amount of PBS buffer was added to cover the cells to prevent the cells from drying out. 2 UVA irradiation was performed for 66.67 min at an irradiation dose of . Then, drugs were administered according to the grouping and drug concentrations of Tables 5 to 7. After 24 h of drug action, the cell culture medium was removed, the cells were washed once with PBS buffer, 1 mL of β-galactosidase staining fixative was added, and the cells were fixed at room temperature for 15 min before the cell fixative was removed. The cells were washed 3 times with PBS buffer, each for 3 minutes; then the PBS buffer was removed, and 1 mL of staining working solution was added to each well (the staining working solution was a mixture of the following liquids: 10 μL β-galactosidase staining solution A, 10 μL β-galactosidase staining solution B, 930 μL β-galactosidase staining solution C, 50 μL X-Gal solution); the culture plate was placed in an incubator at 37°C and incubated overnight. The 96-well plate was sealed with plastic wrap to prevent evaporation, and the plate was observed and photographed under an inverted optical microscope. The results are shown in the figure. Figure 5 The photos were processed with Image J software, the stained senescent cells were counted, and the proportion of stained senescent cells was calculated. The results are shown in Tables 5 to 7.

[0063] Table 5 Number of cells positive for β-galactosidase stained by combined administration of VA and ginsenoside CK

[0064]

[0065] Table 6 Number of cells positive for β-galactosidase staining after combined administration of HPR and ginsenoside CK

[0066]

[0067] Table 7 Number of cells positive for β-galactosidase stained by combined administration of VAPA and ginsenoside CK

[0068]

[0069]

[0070] Depend on Figure 5 As shown in Tables 5 to 7, compared with the blank control group, the β-galactosidase staining in the irradiated control group was significantly deepened; compared with the irradiated control group, the VA, HPR and VAPA combined with ginsenoside CK treatment groups can significantly reduce the β-galactosidase staining level, indicating that VA, HPR and VAPA combined with ginsenoside CK can alleviate the UVA-induced senescence level of HaCaT cells.

[0071] (4) Western Blot method was used to detect the effect of retinoid combination on photoaging cell apoptosis

[0072] According to the above method, the photoaging model was established and the drug was administered. The photoaged HaCaT cells after irradiation and drug administration were incubated for 24 hours, and then the culture medium was discarded, the cells were washed twice with pre-cooled PBS buffer, and strong RIPA lysis buffer containing PMSF was added to lyse the cells on ice for 30 minutes; the lysate was collected in a 1.5mL EP tube using a cell scraper and centrifuged at 12000rpm and 4℃ for 15 minutes, and then the supernatant was collected, and an appropriate amount of 5× protein loading buffer was added to mix and boiled in a boiling water bath for 10 minutes, and stored in a -20℃ refrigerator for standby use; gel was prepared, sampled, and electrophoresed. After the electrophoresis, the membrane was transferred using a Bio-Rad semi-dry transfer instrument. After the transfer, the membrane was blocked with 5% skim milk at room temperature for 1 hour; the primary antibody was diluted with 1×TBST as required and incubated at room temperature for 2 hours. After the primary antibody incubation, the membrane was washed 3 times with 1×TBST buffer, each time for 7 minutes; then the secondary antibody was added and incubated at room temperature for 1 hour; after the incubation, the membrane was washed with 1×TBST buffer, and finally ECL developer was added to the gel imager for photography and development. The results are shown in the figure. Figure 6 Image J software was used to analyze and process WB photos and calculate the relative protein expression. The results are shown in Figure 7 The results show that UVA radiation leads to increased apoptosis of skin keratinocytes, and the retinol or retinol derivative and ginsenoside CK composition provided by the present invention can effectively inhibit apoptosis of photoaged HaCaT cells.

[0073] (5) Western Blot method was used to detect the effect of retinoid combination on collagen in photoaged cells

[0074] Type I collagen is the main component of skin structure, accounting for 75% of skin weight. It has the function of maintaining skin elasticity and tension and is closely related to skin aging. Elastin is the main component of elastic fibers and plays an important role in skin elasticity. The Western Blot method was used to detect the effect of the retinoid composition on photoaging cell collagen. The test steps were the same as the above experiment (2). The results are as follows: Figure 8 Image J software was used to analyze and process WB photos and calculate the relative protein expression. The results are shown in Fig. 9 The results showed that UVA radiation caused downregulation of the expression of type I collagen (CollagenI) and elastin (Elastin) in HaCaT cells, and ginsenoside CK combined with retinol or retinol derivatives could increase the expression of type I collagen and elastin in a concentration gradient-dependent manner. The above results indicate that the combination of retinol and its derivatives with ginsenoside CK can induce increased expression of collagen and elastin in photoaged cells.

[0075] Based on the above experimental results, it can be seen that VA, HPR and VAPA alone cannot promote the proliferation of UVA photoaged cells, but after being co-administered with ginsenoside CK in different mass ratios, they can promote the proliferation activity of HaCaT cells to varying degrees. The mass ratio of retinol to ginsenoside CK is 1:0.02-2, the mass ratio of HPR to ginsenoside CK is 1:0.002-0.5, and the mass ratio of VAPA to ginsenoside CK is 1:0.002-0.5, all of which have the effect of promoting the proliferation of HaCaT cells. Among them, the combined administration of VA and ginsenoside CK at a mass ratio of 1:1, HPR and ginsenoside CK at a mass ratio of 1:0.2, and VAPA and ginsenoside CK at a mass ratio of 1:0.1 has the most significant effect on promoting the proliferation of photoaged HaCaT cells. Moreover, the combined administration of retinol and its derivatives with ginsenoside CK could significantly reduce the number of positive cells stained by β-galactosidase, indicating that VA, HPR and VAPA combined with ginsenoside CK could alleviate the senescence level of UVA-induced aged HaCaT cells. In addition, the combined administration of retinol and its derivatives with ginsenoside CK significantly reduced UVA-induced apoptosis of HaCaT cells by inhibiting the expression of PARP, Caspase-8 and Caspase-9 proteins in HaCaT cells in a concentration-dependent manner. UVA irradiation caused the downregulation of type I collagen and elastin expression in HaCaT cells. The combined treatment of retinol and its derivatives with ginsenoside CK could increase the expression of type I collagen (Collagen I) and elastin (Elastin) in a concentration-dependent manner.

[0076] (6) Using the fluorescent probe DCFH-DA to detect the effect of retinoid composition on reactive oxygen species in photoaged cells

[0077] Reactive oxygen species (ROS) can damage collagen in the dermis and harden it, which is closely related to skin aging. The fluorescent probe DCFH-DA reactive oxygen species detection kit was used to detect the effect of retinoid composition on reactive oxygen species in photoaged cells. The test steps are as follows: HaCaT cells were digested with trypsin and centrifuged; according to the cell growth rate, 2.0×10 5 The cells were plated at a density of 100 cells / well in a six-well plate, 2 mL of cell suspension was added / well, and the culture plate was placed in an incubator (37°C, 5% CO 2) for 24 hours; after the cells adhered, the drugs were administered according to the grouping and drug concentrations in Tables 8 to 10. After 24 hours of drug action, the cell culture medium was removed, DCFH-DA was diluted with serum-free culture medium at 1:1000 to a final concentration of 10 μM / L, and the diluted DCFH-DA was added to the six-well plate, 1 mL / well. Incubate in a cell culture incubator at 37°C for 20 minutes. Then the cells were collected and detected by flow cytometry. The intensity of fluorescence before and after drug stimulation was detected using an excitation wavelength of 488 nm and an emission wavelength of 525 nm. Among them, E1, F1 and G1 were blank control groups, which were not irradiated with UVA or administered; E2, F2 and G2 were reactive oxygen positive control groups, E3, F3 and G3 were empty probe control groups, and E4, F4 and G4 were irradiated control groups; E5 to E7, F5 to F7 and G5 to G7 were drug administration groups, which were irradiated with UVA and administered at the same time.

[0078] Table 8 Effect of combined administration of VA and ginsenoside CK on ROS

[0079]

[0080] Table 9 Effect of combined administration of HPR and ginsenoside CK on ROS

[0081]

[0082] Table 10 Effect of combined administration of VAPA and ginsenoside CK on ROS

[0083]

[0084]

[0085] The results in Tables 8 to 10 show that compared with the empty probe control group (E3, F3 and G3), UVA radiation (E4, F4 and G4) caused a significant increase in ROS in HaCaT cells, and ginsenoside CK combined with retinol or retinol derivatives could significantly reduce the level of reactive oxygen species.

[0086] Example 2

[0087] According to the grouping and administration concentration of each drug in Tables 11 to 13, retinol or its derivatives and ginsenoside CK are added to butanediol, mixed and stirred evenly to obtain a composition solution; mullein exosomes are added to PBS buffer, the mass ratio of mullein exosomes to PBS buffer is 1:10, and after mixing evenly, the composition solution is added dropwise at a rate of 8 mL / min, ultrasonically treated at a power of 300 W for 2 min under ice water bath conditions, and then incubated at room temperature for 12 h, centrifuged at a centrifugal force of 150000 g for 60 min, the supernatant is discarded, the precipitate is taken, and freeze-dried to obtain a retinoid composition preparation. The exosomes of mullein were prepared by the following method: fresh mullein was washed and dried and then added to PBS buffer solution, the mass ratio of the dried mullein to PBS buffer solution was 1:10, the dried mullein was ground at a speed of 1000r / min for 15min, filtered with gauze, and the filtrate was collected; the collected filtrate was centrifuged at a centrifugal force of 2000g for 30min, and the supernatant was taken; the supernatant was centrifuged at a centrifugal force of 150000g for 80min, the supernatant was discarded, the precipitate was taken, and the mullein exosomes were obtained by freeze drying. Then, the cytotoxicity test was performed according to the method in Example 1, wherein H1, I1 and J1 were blank control groups, the blank control groups were not subjected to UVA irradiation and no drug administration; H2, I2 and J2 were irradiation control groups, only UVA irradiation was performed without drug administration; H3~H7, I3~I7 and J3~J7 were drug administration groups, and the drug administration groups were subjected to UVA irradiation and drug administration at the same time.

[0088] Table 11 Effect of combined administration of VA / ginsenoside CK / verbascum exosomes on proliferation of UVA-irradiated HaCaT cells

[0089]

[0090]

[0091] Table 12 Effect of combined administration of HPR / ginsenoside CK / verbascum exosomes on proliferation of UVA-irradiated HaCaT cells

[0092]

[0093] Table 13 Effect of combined administration of VAPA / ginsenoside CK / verbascum exosomes on proliferation of UVA-irradiated HaCaT cells

[0094]

[0095]

[0096] The above experimental results show that VA, HPR and VAPA alone cannot promote the proliferation of UVA photo-aged cells, but can promote the proliferation activity of HaCaT cells to varying degrees after being combined with ginsenoside CK. In addition, when combined with different concentrations of (0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL) of mullein exosomes, it can significantly promote the proliferation activity of photo-aged HaCaT cells and reduce the irritation of retinol and its derivatives.

[0097] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A retinoid composition for reducing irritation and anti-aging, characterized in that: It comprises anti-aging ingredients and ginsenoside CK, wherein the anti-aging ingredients comprise at least one of retinol and retinol derivatives, and the mass ratio of the anti-aging ingredients to the ginsenoside CK is 1:0.001-2.

2. The anti-irritation and anti-aging retinoid composition according to claim 1, characterized in that: The anti-aging ingredient is retinol, and the mass ratio of the retinol to the ginsenoside CK is 1:0.02-2.

3. The anti-irritation and anti-aging retinoid composition according to claim 1, characterized in that: The anti-aging ingredient is retinol, and the mass ratio of the retinol to the ginsenoside CK is 1:0.1-1.

4. The anti-irritation and anti-aging retinoid composition according to claim 1, characterized in that: The retinol derivative includes at least one of hydroxypinacolone retinoic acid ester, retinol palmitate, retinol propionate, retinol retinoic acid ester, retinol linoleate, and retinol acetate.

5. The anti-irritation and anti-aging retinoid composition according to claim 4, characterized in that: The anti-aging ingredient is hydroxypinacolone retinoic acid ester, and the mass ratio of the hydroxypinacolone retinoic acid ester to the ginsenoside CK is 1:0.002-0.

5.

6. The anti-irritation and anti-aging retinoid composition according to claim 4, characterized in that: The anti-aging ingredient is hydroxypinacolone retinoic acid ester, and the mass ratio of the hydroxypinacolone retinoic acid ester to the ginsenoside CK is 1:0.02-0.

2.

7. The anti-irritation and anti-aging retinoid composition according to claim 4, characterized in that: The anti-aging ingredient is retinol palmitate, and the mass ratio of the retinol palmitate to the ginsenoside CK is 1:0.002-0.

5.

8. The anti-irritation and anti-aging retinoid composition according to claim 4, characterized in that: The anti-aging ingredient is retinol palmitate, and the mass ratio of the retinol palmitate to the ginsenoside CK is 1:0.01-0.

1.

9. Use of the retinoid composition according to any one of claims 1 to 8 in the preparation of daily cosmetics.

10. Use of the retinoid composition according to claim 9 in preparing daily cosmetics, characterized in that: The mass percentage of the retinoid composition in daily cosmetics is 0.01% to 10%.

11. Use of the retinoid composition according to claim 9 in preparing daily cosmetics, characterized in that: The daily cosmetics include any one of facial cleanser, shower gel, skin care lotion, skin care cream, essence, eye cream, and facial mask.

12. A retinoid composition preparation, characterized in that: The invention comprises the retinoid composition according to any one of claims 1 to 5 and verbabasin exosomes, wherein the mass ratio of the retinoid composition to the verbabasin exosomes is 1 to 120:

1.

13. The retinoid composition preparation according to claim 12, characterized in that: The mass ratio of the retinoid composition to the verbabasin exosomes is 2 to 15:

1.

14. A method for preparing the retinoid composition preparation according to claim 12, characterized in that: The retinoid composition is dispersed in butanediol to obtain a composition solution; after the verbascosum exosomes and PBS buffer are mixed, the composition solution is added dropwise, and the retinoid composition preparation is obtained through ultrasonic treatment, incubation, centrifugation, and freeze-drying.

15. The method for preparing the retinoid composition preparation according to claim 14, characterized in that: The dropping rate is 1-10 mL / min; the power of the ultrasonic treatment is 200-500 W, and the ultrasonic time is 1-5 min; the incubation temperature is room temperature, and the incubation time is 12-24 h.

16. Use of the retinoid composition preparation according to claim 12 or 13 in the preparation of daily cosmetics.

17. Use of the retinoid composition preparation according to claim 16 in the preparation of daily cosmetics, characterized in that: The daily cosmetics include any one of facial cleanser, shower gel, skin care lotion, skin care cream, essence, eye cream, and facial mask.

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