CN112386639A

CN112386639A - Application of golden camellia juice in preparing blue light resistant composition

Info

Publication number: CN112386639A
Application number: CN202010823616.4A
Authority: CN
Inventors: 林咏翔; 李姿仪
Original assignee: Bayote Biotechnology Shanghai Co ltd
Current assignee: Bayote Biotechnology Shanghai Co ltd
Priority date: 2019-08-14
Filing date: 2020-08-14
Publication date: 2021-02-23
Also published as: CN112386621A; TWI809300B; TW202106320A; TWI747444B; TW202106327A

Abstract

Use of Camellia nitidissima juice for preparing anti-blue light composition, wherein blue light wavelength is 400 nm to 600 nm. The compositions of the present invention have one or more of the following functions: improving the blue light damage resistance of the skin, improving the blue light damage resistance of skin fiber cells, reducing the generation of active oxidation substances of the skin fiber cells and the like.

Description

Application of golden camellia juice in preparing blue light resistant composition

Technical Field

The invention relates to golden camellia juice, in particular to application of the golden camellia juice in preparing a blue light resistant composition.

Background

Camellia nitidissima is evergreen shrub or small tree of Camellia (Camellia) of Theaceae. Leaves with broad lanceolate to oblong shapes alternate. Golden flower is single-grown leaves axilla or near-top-grown, has petal meat, has waxy luster, is cup-shaped, pot-shaped or bowl-shaped when being opened, and has a flower diameter of 6 cm; 9-11 petals; the flowering period is 10-12 months. The golden camellia is an excellent garden ornamental flower.

In the year 1984, the camellia chrysantha is listed as a Chinese primary protection plant, and the leaf part of the camellia chrysantha is also made into east tea which is reputed to be international. The compendium of materia medica also describes that it has the medicinal functions of clearing away heat and toxic material, promoting diuresis and removing dampness and stopping bleeding.

Disclosure of Invention

In taiwan patent publication No. 201929891, the inventors have disclosed the use of camellia nitidissima juice for enhancing anti-glycosylation activity and inhibiting the accumulation of cell fat. However, in order to improve the medicinal value of the camellia nitidissima, the inventor continuously researches and develops other applications of related products of the camellia nitidissima.

In view of the above, the present invention provides a use of camellia nitidissima juice for preparing a composition for resisting blue light, wherein the wavelength of the blue light is 400 nanometers (nm) to 600 nanometers (nm).

In one embodiment, the Camellia nitidissima juice is used for improving the skin resistance to blue light damage.

In one embodiment, Camellia nitidissima juice is used for preventing skin fiber cells from being damaged by blue light.

In one embodiment, Camellia nitidissima juice is used to prevent Reactive Oxygen Species (ROS) from exposing skin fibroblasts to blue light.

In one embodiment, Camellia nitidissima juice is used to reduce the production of active oxidizing substances by skin fibroblasts by more than 50%.

In one embodiment, the Camellia nitidissima juice has an effective concentration of 0.25 mg/mL.

In one embodiment, the Camellia nitidissima juice has a total flavone content of 10 mg/g.

In one embodiment, the Camellia nitidissima juice is extracted from leaves of Camellia nitidissima.

In one embodiment, the Camellia nitidissima juice is in powder form.

In summary, Camellia nitidissima juice according to any embodiment of the present invention can be used for preparing a blue light-resistant composition. In other words, the aforementioned composition has one or more of the following functions: improving the blue light damage resistance of the skin, improving the blue light damage resistance of skin fiber cells, reducing the generation of active oxidation substances of the skin fiber cells and the like.

Drawings

FIG. 1 is a histogram of the results of blue light resistance experiment of Camellia nitidissima juice.

Detailed Description

As used herein, the concentration symbol "wt%" generally refers to weight percent concentration, while the concentration symbol "vol%" generally refers to volume percent concentration. As used herein, "Camellia nitidissima" refers generally to the leaves of Camellia nitidissima.

In some embodiments, the Camellia nitidissima juice refers to a product obtained by grinding, extracting, filtering, concentrating, sterilizing, and drying leaves of Camellia nitidissima (Camellia chrysantha). For example, the leaf of camellia nitidissima is cut up, extracted with an extraction solvent to obtain an initial extract, and then the initial extract is filtered to remove impurities, and then concentrated with the filtered initial extract to obtain a concentrated solution. The concentrated solution is then sterilized and the sterilized concentrated solution is spray dried to a powder. The dried powder is golden camellia juice containing effective components.

In some embodiments, the Camellia nitidissima juice can be a commercially available Camellia sinensis extract or Camellia sinensis extract powder. In some embodiments, the Camellia nitidissima juice is extracted by pulverizing Camellia nitidissima (Camellia chrysanthha) of Guangzhou into fragments of about less than 0.5cm, mixing the pulverized Camellia nitidissima with a solvent at a temperature of 80 ± 10 ℃ in a range of 5-20: extracting for 0.5-2 hours at a volume ratio of 1-5 to obtain a crude juice. Wherein the solvent may be water, alcohols, aqueous alcohols, or combinations thereof. Subsequently, the crude juice was cooled to room temperature, and then the crude juice was centrifuged at 5000rpm for 10 minutes and subjected to desludging filtration, and the supernatant was collected. Thereafter, the supernatant was filtered through a 400 mesh (mesh) sieve to obtain a filtrate, and then the filtrate was concentrated under reduced pressure at 55. + -. 10 ℃ to obtain a concentrated product. And then, carrying out spray drying on the concentrated product to obtain golden camellia juice.

In one embodiment, the Camellia nitidissima juice is in powder form. In one embodiment, the Camellia nitidissima juice is an edible grade powder.

In an embodiment, the use of camellia nitidissima juice for the preparation of a composition for resisting blue light, wherein the blue light wavelength is 400 nanometers (nm) to 600 nanometers (nm).

In one embodiment, the Camellia nitidissima juice is used for improving the skin resistance to blue light damage. In one embodiment, Camellia nitidissima juice is used for preventing skin fiber cells from being damaged by blue light. In one embodiment, Camellia nitidissima juice is used to prevent skin fibroblasts from Reactive Oxidative Species (ROS) generated by blue light irradiation.

In some embodiments, any of the compositions described above can be a pharmaceutical. In other words, the medicine contains the golden camellia juice with effective content.

In some embodiments, the foregoing pharmaceuticals may be manufactured using techniques well known to those skilled in the art into dosage forms suitable for enteral, parenteral (parenterally), oral, or topical (topically) administration.

In some embodiments, the dosage form for enteral or oral administration may be, but is not limited to, a lozenge (tablet), a tablet (troche), a buccal tablet (dosage), a pill (pill), a capsule (capsule), a dispersible powder (dispersible powder) or fine granules (granules), a solution, a suspension (suspension), an emulsion (emulsion), a syrup (syrup), an elixir (elixir), a slurry (syrup), or the like. In some embodiments, the parenteral or topical administration dosage form can be, but is not limited to, an injectable (injection), a sterile powder (sterile powder), an external preparation (external preparation), or the like. In some embodiments, the administration of the injectate can be subcutaneous (subcutaneous), intradermal (intraepithelial injection), or intralesional (intrafocal injection).

In some embodiments, the aforementioned pharmaceutical may comprise a pharmaceutically acceptable carrier (pharmaceutical acceptable carrier) that is widely used in pharmaceutical manufacturing technology. In some embodiments, the pharmaceutically acceptable carrier can be one or more of the following carriers: solvents (solvent), buffers (buffer), emulsifiers (emulsifying), suspending agents (suspending agent), disintegrating agents (disintegrant), disintegrating agents (disintegrating agent), dispersing agents (dispersing agent), binding agents (binding agent), excipients (excipient), stabilizers (stabilizing agent), chelating agents (chelating agent), diluents (diluent), gelling agents (gelling agent), preservatives (preserving), wetting agents (wetting agent), lubricants (lubricating), absorption delaying agents (absorption delaying agent), liposomes (liposome) and the like. The type and amount of carrier selected for use is within the skill of the art in terms of skilled literacy and routine skill. In some embodiments, the solvent as a pharmaceutically acceptable carrier may be water, physiological saline (normal saline), Phosphate Buffered Saline (PBS), or an aqueous solution containing alcohol (aqueous solution).

In some embodiments, any of the foregoing compositions may be an edible composition. In other words, the edible composition comprises a specified content of camellia nitidissima juice. In some embodiments, the aforementioned edible composition may be a food product or food additive (food additive). In some embodiments, the food product may be, but is not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).

In some embodiments, any of the foregoing compositions may be a cosmetic or a care product. In other words, the cosmetic or care product contains a specified content of the camellia nitidissima juice.

In some embodiments, the cosmetic or care product may be any of the following types: lotions, gels, jellies, mud masks, lotions, creams, lipsticks, foundations, pressed powders, honey powders, make-up removers, facial cleansers, shower gels, shampoos, hair tonics, sun blocks, hand creams, nail polishes, perfumes, essences, and facial masks. In some embodiments, the cosmetic or care product may further comprise an external acceptable ingredient, if desired. In some embodiments, the topical acceptable ingredient can be, for example, an emulsifier, a penetration enhancer, a softener, a solvent, an excipient, an antioxidant, or a combination thereof.

An example is as follows: total flavone content test

A methanol solution of 200. mu.g/mL of Rutin (Rutin) was taken as the initial solution (i.e., containing 2000ppm of Rutin). Then, 0. mu.g/mL, 20. mu.g/mL, 40. mu.g/mL, 60. mu.g/mL, 80. mu.g/mL, and 100. mu.g/mL of a standard solution of rutin were prepared according to the following Table I, and 200. mu.L of each concentration of the standard solution was taken out into a glass tube, respectively. 200 mu L of 5% sodium citrate aqueous solution is added into each glass test tube and is uniformly mixed with the standard solution and is kept stand for 6 minutes, then 200 mu L of 10% aluminum nitrate aqueous solution is added and is uniformly mixed and is kept stand for 6 minutes, 2mL of 4% sodium hydroxide aqueous solution is added and is uniformly mixed, and then 1.4mL of water is added and is uniformly mixed to obtain the standard reaction solution. A standard curve was obtained by taking 200. mu.L of the standard reaction solution into a 96-well plate and measuring the absorbance thereof at 500 nm.

Watch 1

Standard solution (μ g/mL)	0	200	400	600	800	1000
							Initial solution (μ L)	0	200	400	600	800	1000
Water (mu L)	1000	800	600	400	200	0

Camellia nitidissima juice powder (purchased from Changsha Ha root Biotechnology Co., Ltd., referred to the composition table comprising 92% Camellia nitidissima juice and 8% maltodextrin) was diluted at a ratio of 1:50 (by weight) to obtain a sample. Samples were taken at 200. mu.L into glass tubes. 200 mu L of 5% sodium citrate aqueous solution is added into each glass test tube and is uniformly mixed with the standard solution and is kept stand for 6 minutes, then 200 mu L of 10% aluminum nitrate aqueous solution is added and is uniformly mixed and is kept stand for 6 minutes, 2mL of 4% sodium hydroxide aqueous solution is added and is uniformly mixed, and then 1.4mL of water is added and is uniformly mixed to obtain the standard reaction solution. 200. mu.L of the standard reaction solution was taken into a 96-well plate, and the absorbance at 500nm was measured.

And then, converting the light absorption value of the reaction solution to be detected into the total flavone content by using a standard curve and an interpolation method. Thus, the content of the total flavone in the obtained golden camellia juice is 10 mg/g.

Example two: test for resistance to blue light damage

Human skin fibroblast CCD-966sk (preservation number BCRC60153) was used for this test. The test uses medium of the type MEM (minimum essential medium) of Earle's balanced salts containing 0.1M non-essential amino acids, 1.5g/L sodium bicarbonate, 0.1M pyruvic acid, and 10% fetal bovine serum (from Gibco) by additional addition.

The sample is prepared by using golden camellia juice powder (purchased from Changsha Ha root biotechnology limited) and a culture medium to obtain a golden camellia extraction solution with the concentration of 0.25mg/mL as a first sample and a golden camellia extraction solution with the concentration of 0.125mg/mL as a second sample.

Fluorescent dye DCFH-DA (purchased from Sigma/SI-D6883-50MG) was formulated in Dimethyl sulfoxide (DMSO) as an active oxidizing substance dye at a concentration of 5MG/ml, which could stain against the active oxidizing substance.

The room temperature for this test is 25. + -. 5 ℃.

First, 2mL of medium was added to each well of a six-well plate, and each well was colonized with 1X 10⁵Human dermal fibroblasts. Next, after culturing at 37 ℃ for 24 hours, the medium was removed. The following groups were subjected to duplicate tests and their rounded average values were taken.

Blank group: 0.625. mu.L of medium per well was added and incubated at 37 ℃ for 1 hour, after which 2. mu.L of the active oxidizing substance stain per well was added and allowed to stain for 15 minutes, and after 15 minutes, incubated in the dark at room temperature for 15 minutes.

Control group: after 0.625. mu.L of the medium was added per well and incubated at 37 ℃ for 1 hour, 2. mu.L of the active oxidizing substance dye was added per well to stain for 15 minutes, and after 15 minutes, the wells were placed in a blue light box at room temperature and irradiated with blue light (mainly at 500 nm) for 15 minutes.

Experiment group one: 0.625. mu.L of the above-mentioned sample I was added per well and incubated at 37 ℃ for 1 hour, after which 2. mu.L of the active oxidizing substance stain was added per well and allowed to stain for 15 minutes, after which it was placed in a blue light box at room temperature for 15 minutes under blue light irradiation.

Experiment group two: 0.625. mu.L of the above sample two was added per well and incubated at 37 ℃ for 1 hour, after which 2. mu.L of the active oxidizing substance stain was added per well and allowed to stain for 15 minutes, after which it was placed in a blue light box at room temperature for 15 minutes under blue light irradiation.

Then, each of the above groups was washed twice with 1-fold concentration of PBS buffer (purchased from Gibco), and then reacted for 5 minutes in the absence of light after adding 200ml of trypsin, respectively. The reacted groups are transferred to a 15ml test tube and centrifuged at 400 Xg for 10 minutes, the supernatant in the test tube is removed, then the cells in the test tube are washed once with PBS buffer solution with the concentration of 1 times, then the test tube groups are centrifuged at 400 Xg for 10 minutes again, the supernatant in each group is removed, then 1ml of PBS buffer solution with the concentration of 1 times is added to precipitate the cells in the test tube, finally the fluorescence signals of each group at the excitation wavelength of 450-490 nm and the radiation wavelength of 510-550 nm are detected by a Flow Cytometer (BD Accuri C6 Plus Flow Cytometer 660517) to quantify the content of the reactive oxygen species in the cells, and the proportion of the number of the cells highly expressed by the reactive oxygen species in the cells to the original number is obtained as the relative production amount of the Reactive Oxygen Species (ROS).

The results are shown in Table II below, where the values in Table II are the relative active oxidizing species production, expressed as% calculated by the algorithm in the flow cytometer.

Watch two

	Relative amount of Reactive Oxidizing Species (ROS) produced
		Blank group	3％
Control group	42％
		Experiment set 1	18％
Experiment group two	27％

In this case, the determination of whether statistically significant differences between the two sample populations are observed using Excel software is shown in FIG. 1 (in the figure, "x" represents p values less than 0.05, p values less than 0.01, and "x" represents p values less than 0.001. when "x" is more, the statistical differences are more significant).

Refer to fig. 1. The human skin fibroblasts in the blank group were not irradiated with blue light, which means that they were produced in a relative amount of 3% of active oxidative substances under normal physiological metabolic conditions. The human skin fiber cells of the control group produced up to 42% of the relative active oxidative species after 15 minutes of blue light exposure. It is known that blue light significantly promotes the production of active oxidizing substances by human skin fibroblasts, so that the human skin can be greatly damaged.

Reference is continued to fig. 1. The human skin fiber cells of the second experimental group produced 27% of the active oxidative species after being treated with the gold camellia sap at a concentration of 0.125 mg/mL. The amount of active oxidized species produced in the second experimental group was reduced by 15% compared to the amount of active oxidized species produced in the control group. The experimental result shows that the golden camellia juice with the concentration of 0.125mg/mL can obviously prevent the generation of active oxidation substances.

Whereas the human skin fibroblasts of the experimental group one produced 18% of the relative active oxidized species after being treated with the camellia sinensis juice at a concentration of 0.25 mg/mL. The active oxidizing species production in the experimental group was reduced by up to 24% compared to the relative active oxidizing species production in the control group. The experimental result shows that the golden camellia juice with the concentration of 0.25mg/mL can obviously prevent the generation of active oxidation substances.

The present invention is capable of other embodiments, and various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Use of Camellia nitidissima juice for the preparation of a composition for the treatment of blue light, wherein the blue light has a wavelength of 400 nm to 600 nm.

2. The use of claim 1, wherein the Camellia nitidissima juice is used to improve the skin's resistance to blue light damage.

3. The use of claim 2, wherein the Camellia nitidissima juice is used to prevent blue light damage to skin fibroblasts.

4. The use of claim 3, wherein the Camellia nitidissima juice is used to prevent Reactive Oxygen Species (ROS) in skin fibroblasts from exposure to blue light.

5. The use of camellia nitidissima juice according to claim 4, wherein the camellia nitidissima juice is used to reduce the production of the active oxidizing species by more than 24% of skin fibroblasts.

6. The use according to any one of claims 1 to 5, wherein the Camellia nitidissima juice is present in an effective concentration of 0.25 mg/mL.

7. Use according to any one of claims 1 to 5, wherein the Camellia nitidissima juice has a total flavone content of 10 mg/g.

8. The use according to claim 1, wherein the Camellia nitidissima juice is obtained by extraction of the leaves of Camellia nitidissima.

9. The use according to claim 1, wherein the Camellia nitidissima juice is in powder form.