TWI747444B - Use of camellia chrysantha extract for improving anti-blue light damage effect - Google Patents

Abstract

A use of Camellia Chrysantha extract for improving anti-blue light damage effect, wherein the wavelength of the blue light is 400nm to 600nm.

Description

Use of golden flower tea extract for preparing anti-blue light composition

The present invention relates to a Camellia sinensis extract, in particular to the use of the Camellia sinensis extract to prepare an anti-blue light composition.

Camellia is an evergreen shrub or small tree in the genus Camellia of the family Theaceae. The leaves are alternately wide-lanceolate to oblong. Golden yellow flowers are solitary leaf axils or near terminal, petals fleshy, waxy luster, cup-shaped, pot-shaped or bowl-shaped when opening, flower diameter can reach 6 cm; petals 9-11; flowering October-December. Camellia is an excellent garden ornamental flower and tree.

In 1984, Camellia sinensis was listed as a first-level protected plant in China, and the leaves of Camellia sinensis were also made into an internationally renowned oriental tea. It is also recorded in the Compendium of Materia Medica that it has the medicinal functions of clearing away heat, detoxifying, diuresis and dampness and hemostasis.

In Taiwan Patent Publication No. 201929891, the inventor has disclosed a use of Camellia tea extract to enhance anti-glycation activity and inhibit cell fat accumulation. However, in order to enhance the medicinal value of Camellia tea, the inventor continues to research and develop other uses of Camellia-related products.

In view of this, the present invention provides a use of a golden camellia extract for preparing an anti-blue light composition, wherein the blue light wavelength is 400 nanometers (nm) to 600 nanometers (nm).

In one embodiment, the golden flower tea extract is used to enhance the skin's ability to resist blue light damage.

In one embodiment, the golden flower tea extract is used to prevent skin fiber cells from being damaged by blue light.

In one embodiment, the golden flower tea extract is used to prevent skin fiber cells from being exposed to reactive oxygen species (ROS) generated by blue light irradiation.

In one embodiment, the golden flower tea extract is used to reduce more than 50% of the active oxidant produced by skin fibroblasts.

In one embodiment, the effective concentration of the golden flower tea extract is 0.25 mg/mL.

In one embodiment, the total flavonoid content of the extract of Camellia sinensis is 10 mg/g.

In one embodiment, the golden flower tea extract is obtained from the leaves of the golden flower tea.

In one embodiment, the golden flower tea extract is in powder form.

In summary, the Camellia tea extract according to any embodiment of the present invention can be used to prepare an anti-blue light composition. In other words, the aforementioned composition has one or more of the following functions: enhancing the ability of the skin to resist blue light damage, enhancing the ability of skin fiber cells to resist blue light damage, reducing the production of active oxidants by skin fiber cells, and the like.

Regarding the concentration symbol "wt%" used in this article usually refers to weight percent concentration, and the concentration symbol "vol%" usually refers to volume percent concentration. About the "golden flower tea" used in this article usually refers to the leaves of the gold flower tea.

In some embodiments, the Camellia chrysantha extract refers to a product obtained by grinding, extracting, filtering, concentrating, sterilizing and drying the leaves of Camellia chrysantha (Camellia chrysantha). For example, the leaves of Camellia sinensis are chopped, extracted with an extraction solvent to obtain a preliminary extract, and then the preliminary extract is filtered to remove impurities, and the filtered preliminary extract is concentrated to obtain a concentrated liquid. Next, the concentrated solution is sterilized, and the sterilized concentrated solution is spray-dried to dry into powder. Here, the dried powder is a camellia extract containing active ingredients.

In some embodiments, the golden camellia extract may be a commercially available camellia extract or a camellia extract powder. In some embodiments, the extraction method of Camellia chrysantha extract is to pulverize Camellia chrysantha (Camellia chrysantha), China, into fragments less than 0.5 cm, and take the pulverized Camellia chrysantha at 80±10°C, and compare the Camellia chrysantha with a solvent The volume ratio of 5-20:1-5 is extracted for 0.5-2 hours to obtain a crude extract. Among them, the solvent may be water, alcohols, water-containing alcohols, or a combination thereof. Next, the crude extract was cooled to room temperature, and then the crude extract was centrifuged at 5000 rpm for 10 minutes, deslagging and filtering, and the supernatant was collected. After that, the supernatant was filtered with a 400 mesh filter to obtain a filtrate, and then the filtrate was concentrated under reduced pressure at 55±10° C. to obtain a concentrated product. Next, the concentrated product is spray-dried to obtain a Camellia extract.

In one embodiment, the golden flower tea extract is in powder form. In one embodiment, the golden flower tea extract is an edible powder.

In one embodiment, the use of the golden flower tea extract for preparing an anti-blue light composition, wherein the blue light wavelength is 400 nanometers (nm) to 600 nanometers (nm).

In one embodiment, the golden flower tea extract is used to enhance the skin's ability to resist blue light damage. In one embodiment, the golden flower tea extract is used to prevent skin fiber cells from being damaged by blue light. In one embodiment, the extract of Camellia sinensis is used to prevent skin fiber cells from being exposed to reactive oxygen species (ROS) generated by blue light irradiation.

In one embodiment, the golden flower tea extract is used to reduce more than 50% of the active oxidants produced by skin fiber cells.

In one embodiment, the effective concentration of the golden flower tea extract is 0.25 mg/mL.

In one embodiment, the total flavonoid content of the extract of Camellia sinensis is 10 mg/g.

In some embodiments, any of the aforementioned compositions may be pharmaceuticals. In other words, this medicine contains an effective content of Camellia sinensis extract.

In some embodiments, the aforementioned medicines can be manufactured to be suitable for enteral, parenterally, oral, or topically using techniques well known to those skilled in the art. Dosage type.

In some embodiments, the dosage form for enteral or oral administration may be, but is not limited to, a tablet, a troche, a lozenge, a pill, and a capsule. , Dispersible powder or granule, solution, suspension, emulsion, syrup, elixir, slurry or similar. In some embodiments, the dosage form for parenteral or topical administration may be, but is not limited to, injection, sterile powder, external preparation, or the like. In some embodiments, the injection method may be subcutaneous injection, intraepidermal injection, intradermal injection, or intralesional injection.

In some embodiments, the aforementioned pharmaceutical products may include a pharmaceutically acceptable carrier that is widely used in pharmaceutical manufacturing technology. In some embodiments, the pharmaceutically acceptable carrier may be one or more of the following carriers: solvent, buffer, emulsifier, suspending agent, decomposing agent ( decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent ), gelling agent, preservative, wetting agent, lubricant, absorption delaying agent, liposome and the like. Regarding the type and quantity of the selected carrier, it falls within the scope of professionalism and routine technology of those who are familiar with this technology. In some embodiments, the solvent as a pharmaceutically acceptable carrier may be water, normal saline (normal saline), phosphate buffered saline (PBS), or aqueous solution containing alcohol (aqueous solution containing alcohol). ).

In some embodiments, any of the aforementioned compositions may be edible compositions. In other words, the edible composition contains a specific content of Camellia sinensis extract. In some embodiments, the aforementioned edible composition may be a food product or a food additive. In some embodiments, the food product may be, but is not limited to: beverages, fermented foods, bakery products, health foods, and dietary supplements.

In some embodiments, any of the aforementioned compositions may be cosmetics or skin care products. In other words, cosmetics or skin care products contain a specific content of Camellia tea extract.

In some embodiments, the aforementioned cosmetics or skin care products can be any of the following types: lotion, gel, jelly film, mud mask, lotion, cream, lipstick, foundation, powder, powder, makeup remover, makeup remover Milk, facial cleanser, body wash, shampoo, hair conditioner, sunscreen, hand cream, nail polish, perfume, essence and facial mask. In some embodiments, the aforementioned cosmetics or skin care products may further include externally acceptable ingredients as needed. In some embodiments, the external product acceptable ingredient may be, for example, an emulsifier, a penetration enhancer, a softener, a solvent, an excipient, an antioxidant, or a combination thereof.

Demonstration example 1: Total flavonoid content test

The 200μg/mL Rutin methanol solution was obtained as the initial solution (that is, containing 2000ppm Rutin). Then, configure the standard solutions of 0μg/mL, 20μg/mL, 40μg/mL, 60μg/mL, 80μg/mL, and 100μg/mL according to Table 1 below, and take 200μL of the standard solutions of each concentration to In a glass test tube. Add 200μL of 5% sodium citrate aqueous solution to each glass test tube and mix it with the standard solution and let it stand for 6 minutes, then add 200μL of 10% aluminum nitrate aqueous solution to mix well and let it stand for 6 minutes, then add 2 mL of 4% After the sodium hydroxide aqueous solution is evenly mixed, add 1.4 mL of water and mix well to obtain a standard reaction solution. Take 200μL of the standard reaction solution into a 96-well plate, and measure its absorbance at 500nm to obtain a standard curve.

Table I Standard solution (μg/mL) 0 200 400 600 800 1000 Initial solution (μL) 0 200 400 600 800 1000 Water (μL) 1000 800 600 400 200 0

Dilute Camellia tea extract powder (purchased from Changsha Hagen Biotechnology Co., Ltd., refer to its ingredient list which includes 92% Camellia tea extract and 8% maltodextrin) at 1:50 (weight ratio) Later as a sample. Take 200 μL of the sample into a glass test tube. Add 200μL of 5% sodium citrate aqueous solution to each glass test tube and mix it with the standard solution and let it stand for 6 minutes, then add 200μL of 10% aluminum nitrate aqueous solution to mix well and let it stand for 6 minutes, then add 2 mL of 4% After the sodium hydroxide aqueous solution is evenly mixed, add 1.4 mL of water and mix well to obtain a standard reaction solution. Take 200μL of the standard reaction solution into a 96-well plate, and measure its absorbance at 500nm.

Then, the absorbance value of the reaction solution to be tested is converted into the total flavonoid content using the standard curve and interpolation method. Here, the total flavonoid content of the extract of Camellia sinensis is 10 mg/g.

Demonstration Example 2: Anti-blue light damage test

This test uses human skin fiber cells CCD-966sk (preservation number BCRC 60153). The medium used in this test is the Earle's balanced salt type that contains 0.1M non-essential amino acid, 1.5g/L sodium bicarbonate, 0.1M pyruvate, and 10% fetal bovine serum (purchased from Gibco) through additional additions MEM (Minimum essential medium) medium.

The sample this time is the Camellia tea extract powder (purchased from Changsha Hagen Biotechnology Co., Ltd.) with a medium configuration of 0.25mg/mL Camellia Camellia extract solution as sample 1 and a Camellia Camellia extract solution with 0.125mg/mL as the sample two.

The fluorescent dye DCFH-DA (purchased from Sigma/SI-D6883-50MG) is formulated with dimethyl sulfoxide (DMSO) into an active oxidizing substance dye with a concentration of 5mg/ml, which can target active oxidizing substances Perform dyeing.

The room temperature referred to in this test is 25±5°C.

First, add 2 mL of medium to each well of the six-well culture plate, and colonize 1×10 ⁵ human skin fibroblasts per well. Next, after culturing at 37°C for 24 hours, the medium was removed. The following groups were subjected to two repeated experiments and the rounded average value was taken.

Blank group: Add 0.625μL of medium to each well and incubate at 37°C for 1 hour, then add 2μL of active oxidizing substance dye to each well to stain for 15 minutes, and then incubate in the dark at room temperature for 15 minutes after 15 minutes minute.

Control group: Add 0.625μL of culture medium to each well and incubate at 37°C for 1 hour, then add 2μL of active oxidizing substance dye to each well to stain for 15 minutes, and then place it in a blue light box at room temperature after 15 minutes. Blue light (mainly 500nm) is irradiated for 15 minutes.

Experimental group 1: Add 0.625μL of the above sample one to each well and incubate at 37℃ for 1 hour, then add 2μL of active oxidizing substance dye to each well to stain for 15 minutes, and then put it in blue light at room temperature after 15 minutes The box was irradiated with blue light for 15 minutes.

Experimental group two: add 0.625μL of the above sample two to each well and incubate at 37°C for 1 hour, then add 2μL of active oxidizing substance dye to each well to stain for 15 minutes, and then place in blue light at room temperature after 15 minutes The box was irradiated with blue light for 15 minutes.

Then, the above groups were washed twice with a 1-fold concentration of PBS buffer (purchased from Gibco), and then 200ml trypsin was added respectively and reacted for 5 minutes in a dark state. After the reaction, each group was transferred to a 15ml test tube and centrifuged at 400×g for 10 minutes. After removing the supernatant in the test tube, the cells in the test tube were washed with 1 times the concentration of PBS buffer once, and then each group was again The test tube was centrifuged at 400×g for 10 minutes. After removing the supernatant of each group, 1ml of 1x PBS buffer was added to precipitate the cells in the test tube. Finally, each group was passed through a flow cytometer (BD Accuri C6 Plus Flow Cytometer 660517) Detect the fluorescence signal at the excitation wavelength of 450~490nm and the emission wavelength of 510~550nm to quantify the content of reactive oxygen species in the cell, and thereby know the ratio of the number of cells with high expression of reactive oxygen species to the number of original cells , As its relative reactive oxygen species (ROS) production.

The experimental results are as follows in Table 2. The values in Table 2 are calculated by the algorithm in the flow cytometer and the relative amount of active oxidants produced is expressed in %.

Table II Relatively active oxidant (ROS) production Blank group 3% Control group 42% Experimental group one 18% Experimental group two 27%

Among them, Excel software is used to perform student t-test to determine whether there is a statistically significant difference between the two sample groups, as shown in Figure 1. The value is less than 0.01, and "***" means that the p-value is less than 0.001. The more "*", the more significant the statistical difference).

Refer to Figure 1. The human skin fiber cells in the blank group were not irradiated with blue light, which means that under normal physiological metabolism, their relative production of active oxidants was 3%. The human skin fiber cells in the control group produced 42% of the relative active oxidants after being irradiated with blue light for 15 minutes. It can be seen that blue light can obviously promote the production of active oxidizing substances in human skin fiber cells, which can cause great damage to human skin.

Continue to refer to Figure 1. After the human skin fibroblasts of experimental group 2 were treated with a 0.125 mg/mL camellia extract, the relative amount of active oxidants produced was 27%. Compared with the relative amount of active oxidizing substances produced in the control group, the amount of active oxidizing substances produced in experimental group two was reduced by 15%. The results of this experiment show that the Camellia extract at a concentration of 0.125mg/mL can significantly prevent the production of active oxidants.

The human skin fibroblasts of experimental group 1 were treated with 0.25mg/mL camellia extract, and the relative active oxidant production was 18%. Compared with the relative amount of active oxidizing substances produced in the control group, the amount of active oxidizing substances produced in the experimental group was reduced by up to 24%. The results of this experiment show that the concentration of 0.25mg/mL Camellia extract can significantly prevent the production of active oxidants.

Although the technical content of the present invention has been disclosed in the preferred embodiments as above, it is not intended to limit the present invention. Anyone who is familiar with this technique and makes some changes and modifications without departing from the spirit of the present invention should be covered by the present invention Therefore, the scope of protection of the present invention shall be subject to the scope of the attached patent application.

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Figure 1 is a bar graph showing the results of the anti-blue light experiment of the extract of Camellia sinensis.

Claims (8)

A use of Camellia tea extract is used to prepare an anti-blue light composition, wherein the wavelength of the blue light is 400 nanometers (nm) to 600 nanometers (nm), and the Camellia tea extract is used to prevent skin fibroblasts Damaged by blue light. The use according to claim 1, wherein the camellia extract is used to enhance the skin's ability to resist blue light damage. The use according to claim 1, wherein the camellia extract is used to prevent skin fiber cells from receiving reactive oxygen species (ROS) generated by blue light irradiation. The use according to claim 3, wherein the golden camellia extract is used to reduce more than 24% of the active oxidative substances produced by skin fibroblasts. The use according to any one of claims 1 to 4, wherein the effective concentration of the golden camellia extract is 0.25 mg/mL. The use according to any one of claims 1 to 4, wherein the total flavonoid content of the camellia extract is 10 mg/g. The use according to claim 1, wherein the golden flower tea extract is obtained by extracting the leaves of the golden flower tea. The use according to claim 1, wherein the camellia extract is in powder form.

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