CN114948816A - A kind of sunscreen and anti-photoaging traditional Chinese medicine composition and application thereof - Google Patents

Abstract

The invention discloses a sun-screening anti-photoaging traditional Chinese medicine composition and application thereof, wherein the sun-screening anti-photoaging traditional Chinese medicine composition comprises effective components and DMSO (dimethylsulfoxide), wherein the effective components comprise a methanol reflux extract of Hibiscus taiwanensis, a 95% ethanol extract of Trillium fortunei and a distilled water extract of gardenia according to a mass ratio of 4: 3, and the concentration of the effective components in the DMSO is 4-8 mg/mL. The invention has the effects of sun screening and anti-photoaging, can be used in cosmetics to achieve the effects of sun screening and anti-aging, and is particularly suitable for rough skin, wrinkle increase, sallow complexion and the like caused by sunlight irradiation; the invention is purely natural, has no stimulation, safety and no toxic and side effects.

Description

A kind of sunscreen and anti-photoaging traditional Chinese medicine composition and application thereof

technical field

The invention belongs to the technical field of cosmetics, and in particular relates to a sunscreen and anti-photoaging traditional Chinese medicine composition and application thereof.

Background technique

Skin aging is a complex biological process, which can be divided into two types: endogenous aging and exogenous aging. Endogenous aging is skin aging that occurs over time and is genetically dependent. Exogenous aging is caused by external environmental factors, among which UV radiation is the main factor, accounting for 80% of the external influence, so exogenous aging is also called photoaging.

The most striking clinical features of photoaging are dry skin, saggy nodules, increased fine lines, loss of elasticity, and hyperpigmentation. Histologically, the photoaging skin has increased melanocytes, hyperpigmentation, marked reduction of Langerhans cells, thickened dermis, marked reduction of collagen fibers, but a large increase in elastic fibers and degeneration. Photoaging at the cellular level is mainly characterized by excessive accumulation of reactive oxygen species (ROS) and changes in extracellular matrix components.

Studies have found that 80% to 90% of skin aging is caused by exogenous factors, among which UV radiation is the main reason for accelerating skin aging. Skin photoaging involves a variety of signal molecules and multiple signal transduction pathways. The molecular mechanisms mainly include DNA damage, oxidative stress, inflammatory response, collagen structure changes and apoptosis. The mechanism of UV-induced photoaging can be found in Open Science (source). Service) Identification Code (OSID). Traditional Chinese medicine is rich in protein and various amino acids, lipids, polysaccharides, flavonoids, vitamins, organic acids, alkaloids, saponins and other nutrients. Its mechanism of action includes repairing DNA damage, anti-oxidation, anti-inflammatory, and skin repair Barrier function and anti-apoptosis, Chinese medicine plays an important role in preventing and delaying skin photoaging.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a sunscreen and anti-photoaging traditional Chinese medicine composition.

Another object of the present invention is to provide the application of the above sunscreen and anti-photoaging traditional Chinese medicine composition.

The technical scheme of the present invention is as follows:

A sunscreen and anti-photoaging traditional Chinese medicine composition is composed of active ingredients and DMSO, wherein the active ingredients are the methanol reflux extract of Taiwan hibiscus, the 95% ethanol extract of Trillium, and the distilled water extract of Gardenia in a ratio of 4:3:3 The composition of the mass ratio, the concentration of active ingredients in DMSO is 4-8mg/mL.

In a preferred embodiment of the present invention, the preparation method of the methanol reflux extract of Hibiscus taiwana includes: taking air-dried rhizomes of Hibiscus taiwana and grinding into powder, extracting with methanol under reflux, and then concentrating under reduced pressure to combine the extracts, A dark brown syrup is obtained, and after concentration, a paste is obtained, which is the methanol reflux extract of Taiwan hibiscus.

In a preferred embodiment of the present invention, the preparation method of the trillium 95% ethanol extract comprises: taking the roots of the trillium, drying in a constant temperature oven, pulverizing with a traditional Chinese medicine pulverizer and passing through a 60-mesh sieve, and using a 95 % ethanol is fully soaked, extracted by a multi-purpose forced percolation tank, repeated several times, the extract is concentrated under reduced pressure, and freeze-dried to obtain the trillium 95% ethanol extract.

In a preferred embodiment of the present invention, the preparation method of the Gardenia jasminoides distilled water extract comprises: accurately weighing the dried gardenia fruit powder, passing through a 60-mesh sieve, dissolving the sieved powder in distilled water, and performing condensation and evaporation after stirring, The liquid is collected and filtered to obtain the first filtrate; the first filtrate is repeatedly centrifuged to collect the supernatant; the supernatant is subjected to suction filtration and the second filtrate is retained, and the second filtrate is subjected to rotary evaporation at 65-75 ° C to obtain viscous liquid; freeze-dry the viscous liquid to obtain dry powder, which is the gardenia distilled water extract.

Application of the above sunscreen and anti-photoaging traditional Chinese medicine composition in the preparation of cosmetics.

In a preferred embodiment of the present invention, the content of the sunscreen and anti-photoaging traditional Chinese medicine composition in the cosmetic is 0.05-10 wt%.

Further preferably, the content of the sunscreen and anti-photoaging traditional Chinese medicine composition in the cosmetic is 0.1-10wt%.

A cosmetic comprising the above sunscreen and anti-photoaging traditional Chinese medicine composition.

In a preferred embodiment of the present invention, the content of the sunscreen and anti-photoaging traditional Chinese medicine composition is 0.05-10 wt%.

Further preferably, the content of the sunscreen and anti-photoaging traditional Chinese medicine composition is 0.1-10wt%.

The beneficial effects of the present invention are:

1. The present invention has sunscreen and anti-photoaging effects, and can be used in cosmetics to achieve sunscreen and anti-aging effects, and is especially suitable for rough skin, increased wrinkles, chlorosis, etc. caused by sunlight.

2. The present invention is pure natural, non-stimulating, safe and non-toxic and side effects.

Description of drawings

1 is a graph showing the detection results of the ability of the sunscreen and anti-photoaging traditional Chinese medicine composition to scavenge DPPH free radicals in Example 5 of the present invention.

2 is a graph showing the results of the detection of the cytotoxicity of the sunscreen and anti-photoaging traditional Chinese medicine composition on HaCaT in Example 5 of the present invention.

3 is a graph showing the detection results of the sunscreen and anti-photoaging traditional Chinese medicine composition in Example 5 of the present invention on the damage and repair activity of UVB-induced HaCaT cells.

4 is a graph showing the effect of the sunscreen and anti-photoaging traditional Chinese medicine composition in Example 5 of the present invention on the ROS content in photoaging HaCaT cells.

Figure 5 is a graph showing the effect of the sunscreen and anti-photoaging traditional Chinese medicine composition in Example 5 of the present invention on the mRNA levels of type I collagen, MMP1 and TIMP1 in HaCaT cells.

6 is a graph showing the effect of the sunscreen and anti-photoaging traditional Chinese medicine composition at different concentrations on the hemolysis rate of hemoglobin in red blood cells in Example 5 of the present invention.

Detailed ways

The technical solutions of the present invention will be further illustrated and described below through specific embodiments in conjunction with the accompanying drawings.

Example 1 Preparation of the methanol reflux extract of Hibiscus Taiwan

The air-dried rhizomes (20kg) of Taiwan hibiscus were ground into powder and extracted 6 times with methanol under reflux. The combined extracts were concentrated under reduced pressure to give a dark brown syrup (850 g) which after concentration gave a paste (400 g).

Example 2 Preparation of Trillium 95% ethanol extract

Trillium tubers (6.4kg) were taken and dried in a constant temperature oven at 48°C, pulverized with a traditional Chinese medicine pulverizer and passed through a 60-mesh sieve, soaked in 95% ethanol for several hours, and extracted using a multi-purpose forced percolation tank. The extract was concentrated under pressure and freeze-dried to obtain an ethanolic trillium extract (2.4 kg).

Example 3 Preparation of Gardenia Distilled Water Extract

Accurately weigh 200g of dried gardenia fruit powder, sieve it with a 0.6mm sieve, weigh 5g of the sieved powder and dissolve it in 250mL of distilled water, stir for 10min, then condense and evaporate for 2h, collect the liquid and filter. The filtrate was centrifuged at 3000r/min for 10min, the supernatant was collected, and the centrifugation was repeated twice; the supernatant was suction filtered and the filtrate was retained, and the filtrate was evaporated in a rotary evaporator at a temperature of 70°C for 30min to obtain a viscous liquid, which was put Put it into a glass plate, pre-freeze it in a -20°C refrigerator overnight, then freeze-dry it for 24 hours, and grind it into powder with a mortar to obtain dry powder (2 g).

Example 4 Preparation of sunscreen and anti-photoaging traditional Chinese medicine composition

According to the mass ratio of 4:3:3, take the methanol reflux extract of Taiwan hibiscus, 95% ethanol extract of Trillium and gardenia distilled water prepared in Examples 1, 2 and 3, respectively, extract and mix to form the active ingredients, add DMSO It is fully dissolved so that the final concentration of the active ingredient is 5 mg/mL, and the sunscreen and anti-photoaging traditional Chinese medicine composition is obtained.

Example 5 Effect experiment of sunscreen and anti-photoaging traditional Chinese medicine composition

Study on the ability of A to scavenge DPPH free radicals

The free radical scavenging activity of peptides was detected by using DPPH free radical scavenging ability detection kit (Biyuntian, BC4750). Take 500 μL of the second reagent solution, thaw on ice, add 2625 μL of the first reagent solution (absolute ethanol), mix well, and put it on ice for use. Get the extract provided by the 1mL test kit and add it to the brown glass tube of Reagent 3. After fully mixing, store 50 μL of each tube in aliquots and place at -20°C; Take out the sunscreen and anti-photoaging Chinese medicine composition from the -20°C refrigerator. Stock solution, add DMSO to dilute, put on ice for use and prepare solvent for future use. Preheat the spectrophotometer for more than 30min, adjust the wavelength to 515nm, and zero in anhydrous ethanol. The 10 mg/mL vitamin C solution (reagent 3) was prepared into a vitamin C solution greater than 0.3 mg/mL with the extract for use (add 1.2 mL of the extract to the storage centrifuge tube and mix well). The sunscreen and anti-photoaging traditional Chinese medicine composition solution, DMSO, extract and working solution were added to the 1.5mL EP tube respectively, so that the final concentrations of the sunscreen and anti-photoaging traditional Chinese medicine composition were 62.5 μM, 125 μM, 250 μM, 500 μM and 1000 μM. After mixing, let it stand for 30min at room temperature in the dark, and the absorbance at 515nm. The absorbance values of blank tube, positive control tube, control tube and assay tube were recorded as A blank, A positive control, A control and A determination, respectively. The blank tube only needs to be tested 1-2 times. The instrument is turned on to warm up and adjust the parameters. The instrument was used for detection, the detection data were recorded and analyzed, and the results were obtained as shown in Figure 1. The sunscreen and anti-photoaging traditional Chinese medicine composition has strong free radical scavenging activity and is concentration-dependent.

Determination of cytotoxicity of sunscreen and anti-photoaging traditional Chinese medicine composition on HaCaT by B CCK-8 method

HaCaT cells were seeded into a 96-well plate (about 4×10 ⁴ /well), and after 24 hours, the cells were completely adherent, the medium was removed, and 100 μL of serum-free medium containing different concentrations of sunscreen and anti-photoaging Chinese medicine compositions was added, and after 24 hours , 10 μL of CCK-8 reagent was added to the 96-well plate, and after 2 h, the OD value was detected at a wavelength of 450 nm. The results are shown in Figure 2. The sunscreen and anti-photoaging Chinese medicine composition has no toxicity to HaCaT cells when the concentration is 0-40 μg/mL, and shows weak cytotoxicity when the concentration reaches 80 μg/mL.

Detection of sunscreen and anti-photoaging traditional Chinese medicine composition on UVB-induced damage and repair of HaCaT cells

HaCaT cells were seeded into a 96-well plate (about 4×10 ⁴ /well), and after 24 hours, the cells were completely adhered, the medium was removed, and 100 μL of serum-free medium containing different concentrations of sunscreen and anti-photoaging Chinese medicine compositions was added for pretreatment for 24 hours. , and then washed 3 times with PBS, and irradiated the HaCaT cells with a certain irradiation dose using a UVB ultraviolet irradiator. After the HaCaT cells were irradiated, the fresh medium with 1% FBS was replaced. After 24 hours, 10 μL of CCK-8 reagent was added to the 96-well plate. After 2 hours, the OD value was detected at a wavelength of 450 nm. The results are shown in Figure 3. It can be seen that the sunscreen and anti-photoaging traditional Chinese medicine composition has a significant protective effect on the photodamage of HaCaT cells when the concentration is 0-40 μg/mL, and the protective effect is when the concentration is 80 μg/mL. weaken.

DEffect of sunscreen and anti-photoaging traditional Chinese medicine composition on ROS content in photoaging HaCaT cells

The production of ROS in HaCaT cells after UVB irradiation was determined by fluorescence microscopy using DCFH2-DA. First, HaCaT cells ( ^1.5 x 105/well) were cultured in medium supplemented with 10% FBS. Then, fresh medium was replaced when 80% cell density was reached. After pretreatment with a certain concentration of sunscreen and anti-photoaging Chinese medicine composition and UVB irradiation, cells were washed with PBS, incubated with 10 μM DCFH2-DA in PBS at 37°C for 30 min, and then washed with PBS buffer. Finally, fluorescence was measured under 485nm excitation and 535nm emission recording spectrophotometer. The results are shown in Figure 4. The sunscreen and anti-photoaging traditional Chinese medicine composition can significantly reduce the content of ROS in photoaging HaCaT cells at a concentration of 0-40 μg/mL. .

E real-time quantitative PCR

RNA was extracted from HaCaT cells pretreated with 40 μg/mL sunscreen and anti-photoaging TCM composition and induced by UVB using the RNeasy mini kit (Qiagen, Pudong, Shanghai, China), followed by GoScript reverse transcription cocktail (Promega, WI) 500ng RNA per sample was reverse transcribed. EVA Green Supermix (Bio-Rad, CA) was then used for real-time quantitative PCR and β-actin was used as an internal control. Primers for MMP1, collagen type I and TIMP1 were designed with Primer5 as shown in Table 1. The 2 ^-ΔCt method was used to quantify relative gene expression.

Table 1

The results are shown in Figure 5. The 40 μg/mL sunscreen and anti-photoaging traditional Chinese medicine composition can partially reverse the decrease in the mRNA expression of type I collagen and TIMP1 and the up-regulation of the mRNA expression of MMP1 in HaCaT cells induced by UVB.

F red blood cell hemolysis test

7 mL of blood was collected from the heart of healthy rabbits, and 1 mL of 2% potassium oxalate solution was added to prepare fresh anticoagulated rabbit blood, diluted with 10 mL of PBS, and stored in a refrigerator at 4°C for later use. Prepare sample series solutions. 8 test doses are set up for each sample, and it must be ensured that the diluted sample suspensions are thoroughly mixed. Determination of hemolysis and protein denaturation: the sample group has 2 test tubes with stoppers for each concentration, and each tube is filled with 10 mL of sample suspension; the negative control group has 2 test tubes with stoppers, and each tube is added with 10 mL of PBS; the positive control group has 2 test tubes with stoppers, each tube is filled with PBS Add 10 mL of distilled water. Add 0.2 mL of diluted rabbit blood to each test tube, mix gently, and incubate at 37°C for 30 min. After the end, centrifuge for 10min, 2000rmin, suck the supernatant and transfer it into a cuvette, and measure its absorbance (A) value at 540nm, 560nm and 575nm with a spectrophotometer. Calculate the hemolysis rate (%) according to the formula ①, and calculate the hemoglobin denaturation index (DI, %) according to the formula ②. R1 is the A ₅₇₅ /A ₅₄₀ value of the centrifugal supernatant of the blank control distilled water; R2 is the A ₅₇₅ /A ₅₄₀ value of the sample centrifugal supernatant; R3 is the A ₅₇₅ /A ₅₄₀ value of the positive control SDS centrifugal supernatant. The main observation indicators were the sample concentration (HD ₅₀ ), protein denaturation index (DI), and the ratio of HD ₅₀ to denaturation index DI (IP=HD ₅₀ /DI) when 50% of erythrocytes were hemolyzed.

The results are shown in Figure 6, the concentration of the sunscreen and anti-photoaging traditional Chinese medicine composition is insoluble in hemolysis below 100 μg/mL.

The above are only the preferred embodiments of the present invention, so the scope of implementation of the present invention cannot be limited accordingly, that is, equivalent changes and modifications made according to the patent scope of the present invention and the contents of the description should still be covered by the present invention. In the range.

Claims (10)

1. A sun-screening and anti-photoaging traditional Chinese medicine composition is characterized in that: the composition comprises effective components and DMSO, wherein the effective components comprise methanol reflux extract of Hibiscus taiwanensis, 95% ethanol extract of Trillium japonicum and distilled water extract of Gardenia jasminoides Ellis at a mass ratio of 4: 3, and the concentration of the effective components in DMSO is 4-8 mg/mL.

2. The sun-screening anti-photoaging traditional Chinese medicine composition of claim 1, wherein: the preparation method of the methanol reflux extract of the mountain Taiwan hibiscus comprises the following steps: grinding the dried rhizomes of the Taiwan mountain hibiscus into powder, performing reflux extraction by using methanol, performing reduced pressure concentration on the obtained product, combining extracts to obtain dark brown syrup, and concentrating the dark brown syrup to obtain paste, namely the methanol reflux extract of the Taiwan mountain hibiscus.

3. The sun-screening anti-photoaging traditional Chinese medicine composition of claim 1, wherein: the preparation method of the Trillium fortunei 95% ethanol extract comprises the following steps: drying the roots of the Trillium wilfordii in a constant-temperature oven, crushing the roots by using a traditional Chinese medicine crusher, sieving the crushed roots by using a 60-mesh sieve, fully soaking the roots in 95% ethanol, extracting the roots by using a multi-purpose forced percolation tank repeatedly, concentrating an extracting solution under reduced pressure, and freeze-drying the extracting solution to obtain the 95% ethanol extract of the Trillium wilfordii.

4. The sun-screening anti-photoaging traditional Chinese medicine composition of claim 1, wherein: the preparation method of the distilled water extract of gardenia comprises the following steps: accurately weighing dried gardenia fruit powder, sieving with a 60-mesh sieve, dissolving the sieved powder in distilled water, stirring, condensing and evaporating, collecting liquid, and filtering to obtain a first filtrate; centrifuging the first filtrate repeatedly to collect supernatant; carrying out suction filtration on the supernatant and reserving a second filtrate, and carrying out rotary evaporation on the second filtrate at 65-75 ℃ to obtain viscous liquid; freeze drying the viscous liquid to obtain dry powder, namely the distilled water extract of cape jasmine.

5. Use of the sunscreen anti-photoaging traditional Chinese medicine composition according to any one of claims 1 to 4 in the preparation of cosmetics.

6. The use of claim 5, wherein: the content of the sun-screening and anti-photoaging traditional Chinese medicine composition in the cosmetics is 0.05-10 wt%.

7. The use of claim 6, wherein: the content of the sun-screening and anti-photoaging traditional Chinese medicine composition in the cosmetics is 0.1-10 wt%.

8. A cosmetic characterized by: a sunscreen anti-photoaging Chinese medicinal composition comprising the composition as claimed in any one of claims 1 to 4.

9. The use of claim 8, wherein: the content of the sun-screening and anti-photoaging traditional Chinese medicine composition is 0.05-10 wt%.

10. The use of claim 9, wherein: the content of the sun-screening and anti-photoaging traditional Chinese medicine composition is 0.1-10 wt%.

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