Anti-blue light composition and application thereof

Abstract

The invention provides an anti-blue light composition and application thereof. The composition disclosed by the invention comprises a hovenia dulcis thunb extract and a nitraria sibirica pall fruit extract. The composition disclosed by the invention can reduce melanin generated by blue light induction by inhibiting tyrosinase activity in B16 cells so as to protect skin from pigmentation caused by blue light irradiation injury.

Description

Technical Field

[0001] The present disclosure relates to the technical field of functional compositions, and in particular to a composition having a protective effect on blue light-induced melanocyte damage and an application thereof. Background Art

[0002] Photoaging is an important factor that causes a series of skin problems. In the past, we paid more attention to the aging damage caused by ultraviolet rays to the skin, but in fact, visible light has stronger skin penetration than ultraviolet rays, and the damage to the skin will be greater. Blue light has the shortest wavelength and the highest energy among all visible light, and exists in natural light and artificial light. Blue light often appears in the light emitted by computer screens, e-books, tablets, smart phones, fluorescent lamps, incandescent lamps, and LEDs in our daily lives. LEDs and OLEDs have unique blue light peaks. Compared with ultraviolet rays, blue light has a longer wavelength and lower energy. Blue light can penetrate the skin layer more deeply and reach the dermis of the skin, so the skin damage caused will be more thorough, such as pigmentation, wrinkles, etc. However, now the public domain research is more about the damage of ultraviolet rays to the skin, and less attention is paid to the skin damage caused by blue light. Combined with the high frequency of exposure to blue light in people's daily work and life, it is worth our attention to how to protect the skin from blue light damage. Summary of the invention

[0003] In order to solve the problems existing in the prior art, the present invention provides a composition having a protective effect on melanocyte damage induced by blue light and its application. Specifically,

[0004] A first aspect of the present invention provides a composition comprising a Hovenia dulcis fruit extract and a Nitraria tangutorum fruit extract.

[0005] In certain embodiments, the composition comprises Hovenia dulcis L. extract and Nitraria tangutorum fructus extract as active ingredients.

[0006] In certain embodiments, the Hovenia dulcis fruit extract is Hovenia dulcis fruit powder.

[0007] In certain embodiments, the content of dihydromyricetin in the Hovenia dulcis extract is ≥ 0.5%.

[0008] In certain embodiments, the Hovenia dulcis powder is a powder obtained by drying a water extract or an alcohol extract of Hovenia dulcis.

[0009] In certain embodiments, the Hovenia dulcis powder is obtained by the following preparation method, which comprises the steps of heating water extraction or alcohol extraction, filtering, concentrating, filtering, drying, pulverizing, and optionally sieving and mixing.

[0010] In certain embodiments, the Nitraria tangutorum fruit extract is Nitraria tangutorum fruit powder.

[0011] In certain embodiments, the Nitraria tangutorum fruit powder is a powder obtained by drying a water extract or an alcohol extract of Nitraria tangutorum fruit.

[0012] In certain embodiments, the Nitraria tangutorum fruit powder is obtained by the following preparation method, which comprises the steps of water extraction or alcohol extraction, concentration, drying, and optionally crushing and sieving.

[0013] In certain embodiments, the weight portion of the Hovenia dulcis extract in the composition is 5-50, and the weight portion of the Nitraria tangutorum fructus extract is 3-10.

[0014] In certain embodiments, the weight percentage of the Hovenia dulcis extract in the composition is 20-50 parts by weight, and the weight percentage of the Nitraria tangutorum fructus extract is 3-5 parts by weight.

[0015] The second aspect of the present invention provides a product comprising the composition described in the first aspect of the present invention.

[0016] In certain embodiments, the product is selected from one or more of a beverage, a tablet, a soft capsule, a hard capsule, a powder, a granule, a candy or a jelly.

[0017] The third aspect of the present invention provides use of the composition described in the first aspect of the present invention in the preparation of food, cosmetics or medicines.

[0018] In certain embodiments, the food, cosmetic or medicine is in a form selected from the group consisting of: one or more of a drink, a tablet, a soft capsule, a hard capsule, a powder, a candy or a jelly.

[0019] In certain embodiments, the food, cosmetic or medicine is used to prevent, inhibit, reduce or eliminate skin pigmentation, or to improve melasma.

[0020] The fourth aspect of the present invention provides the use of the composition described in the first aspect of the present invention or the product described in the second aspect of the present invention in preventing, inhibiting, alleviating or eliminating skin pigmentation or in improving chloasma; preferably, the use is for non-therapeutic purposes.

[0021] In certain embodiments, the skin pigmentation is induced by blue light.

[0022] In certain embodiments, the composition or product inhibits tyrosinase activity and reduces blue light-induced melanin production.

[0023] The fifth aspect of the present invention provides a method for inhibiting tyrosinase activity or reducing blue light-induced melanin production, comprising the step of contacting the composition of the first aspect of the present invention or the product of the second aspect of the present invention with cells.

[0024] In certain embodiments, the cell is a skin cell, preferably a melanocyte.

[0025] In certain embodiments, the method of inhibiting tyrosinase activity or reducing blue light-induced melanin production is for non-therapeutic purposes.

[0026] The present invention has the following advantages over the prior art:

[0027] Provided is a composition having a protective effect on melanocyte damage induced by blue light, which can protect the skin from pigmentation caused by blue light damage by inhibiting tyrosinase activity in B16 cells and reducing melanin generated by blue light. Compared with a single material, the composition has a significant synergistic effect on improving pigmentation induced by blue light.

[0028] The present invention optimizes the ratio of Hovenia dulcis powder and Nitraria tangutorum fruit powder. When the Hovenia dulcis powder is 5-50 parts and the Nitraria tangutorum fruit powder is 3-10 parts, in vitro experiments verify that the powder can protect the skin from pigmentation caused by blue light damage by inhibiting tyrosinase activity and reducing melanin production. Among them, when the Hovenia dulcis powder is 20-50 parts and the Nitraria tangutorum fruit powder is 3-5 parts, the effect of inhibiting tyrosinase activity and reducing melanin production is more significant.

[0029] All ingredients in the composition of the present invention are common food raw materials permitted by laws and regulations and are highly safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The inhibition rate (%) of tyrosinase in the experimental group compared with the model group is shown. Among them, M is the model group and C is the blank control group. The model group is a blue light-damaged model tyrosinase-high-expressing B16 cell irradiated with a 100 Lux blue light source for 10 minutes, and the blank control group is a normal B16 cell wrapped in tin foil to avoid light.

[0031] Figure 2 The relative content of melanin is shown. Among them, M is the model group and C is the blank control group. The model group is a blue light-damaged model of melanin-high-expressing B16 cells irradiated with a 100 Lux blue light source for 10 minutes, and the blank control group is a normal B16 cell wrapped in tin foil to avoid light. DETAILED DESCRIPTION

[0032] The following definitions are provided to facilitate understanding of the present invention by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs. Preferred materials and methods are described herein, but any methods and materials similar to or equivalent to those described herein may be used in the practice of the present invention's tests. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to be limiting.

[0033] Definition of terms

[0034] Unless otherwise indicated or defined, all terms used have the usual meaning in the art, which will be understood by those skilled in the art. In addition, unless otherwise specified, all methods, steps, techniques and operations not specifically described in detail can and have been performed in a manner known per se, which will be understood by those skilled in the art.

[0035] As used herein, the term "Hovenia dulcis" refers to the mature seeds of Hovenia dulcis Thunnb., a plant of the genus Hovenia in the family Rhamnaceae. The known pharmacological effects of Hovenia dulcis include central nervous system inhibitory effects, hypotensive effects, and anti-lipid peroxidation effects.

[0036] As used herein, the term "white thorn fruit" refers to "Qaidam white thorn fruit", which is the fruit of the white thorn of the shrub Nitraria tangutorum Bobrov of the Nitraria family. White thorn (Nitraria tangutorum Bobrov) is a shrub of the Nitraria family, which can reach 2 meters in height. It has many branches, with curved branches and needle-like thorns at the tip. The young branches are broadly oblanceolate, oblong-shaped and spoon-shaped. The flowers are densely white, and the petals and ovaries are glabrous; the flowering period is May-June, and the fruiting period is July-August. Because its branches are white and have thorns at the tip, it is named white thorn.

[0037] Whitethorn fruit is the fruit of the shrub plant Whitethorn of the genus Whitethorn of the family Nitraria. The drupe is ovate, sometimes oval, dark red when ripe, the juice is rose-colored, sweet and sour, with the aroma of grapes and cherries, and is also called "desert cherry". The fruit of Qaidam whitethorn is warm in nature and sweet in taste. It has the effects of strengthening the spleen and digestion, nourishing and calming the mind. The pulp and core of the fruit of Qaidam whitethorn fruit are rich in flavonoids. Flavonoids and their derivatives are effective drugs for anti-inflammatory, cough and expectorant, lowering blood pressure, dilating coronary blood vessels, treating varicose veins, phlebitis, burns and anti-cancer. The nutritional value of the fruit of Qaidam whitethorn is high, rich in vitamin C, and is a good raw material for the beverage industry.

[0038] The term "about" indicates that the value or range of values ​​may vary to an extent that is considered reasonable by one of ordinary skill in the art while still describing a particular property. Specifically, when the term "approximately" is used in this context, it indicates that the value or range of values ​​may vary by 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% of the value or range of values.

[0039] Herein, percentages or % are percentages by weight unless otherwise stated. DETAILED DESCRIPTION OF THE INVENTION

[0041] Composition with protective effect on blue light-induced melanocyte damage

[0042] The invention provides a composition having a protective effect on blue light-induced melanocyte damage, the composition comprising a Hovenia dulcis fruit extract and a Nitraria tangutorum fruit extract.

[0043] In certain embodiments, the composition comprises Hovenia dulcis L. extract and Nitraria tangutorum fructus extract as active ingredients.

[0044] In certain embodiments, the Hovenia dulcis fruit extract is Hovenia dulcis fruit powder.

[0045] In certain embodiments, the Hovenia dulcis powder is prepared by a water extraction method.

[0046] In certain embodiments, the preparation of the Hovenia dulcis powder comprises the steps of heating water extraction, filtering, concentrating, filtering, drying, pulverizing, and optionally sieving and mixing.

[0047] In certain embodiments, in the water extraction step, the mass ratio of Hovenia dulcis to water is 1:(6-12), for example 1:8; the heating temperature is 80-110° C., for example 100° C.; and the extraction time is 1-3 h, for example 2 h.

[0048] In certain embodiments, the filtration step after water extraction uses a filter; preferably, the filter has a mesh size of 200-400, such as 400 mesh.

[0049] In certain embodiments, the filtration step after concentration uses a ceramic membrane; preferably, the pore size of the ceramic membrane is 100-800 nm, such as 100 nm.

[0050] In some embodiments, the concentration step is performed by vacuum concentration; preferably, the concentration vacuum degree is -0.06 to -0.1 MPa, the temperature is about 60-70°C, and the relative density of the concentrate is about 1.10 g-1.14 / cm 3 .

[0051] In certain embodiments, the drying is performed by spray drying.

[0052] In certain embodiments, the sieving step uses an 80 mesh sieve.

[0053] In certain embodiments, the Nitraria tangutorum fruit extract is Nitraria tangutorum fruit powder.

[0054] In certain embodiments, the Nitraria tangutorum fruit powder is prepared by water extraction.

[0055] In certain embodiments, the preparation of the Nitraria tangutorum fruit powder comprises water extraction, concentration, drying, pulverization, and sieving.

[0056] In certain embodiments, the two leachates are combined.

[0057] In certain embodiments, the extract is concentrated by cryogenic vacuum.

[0058] In certain embodiments, the concentrated extract is dried.

[0059] In certain embodiments, the dried drug is crushed and sieved; preferably, sieved through a 40-150 mesh sieve.

[0060] In certain embodiments, the weight proportion of the Hovenia dulcis extract in the composition is 3-60, preferably 5-55, more preferably 5-50, for example about 3, about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, about 55, about 56, about 57, about 58, about 59, about 60.

[0061] In certain embodiments, the weight proportion of the Nitraria tangutorum extract in the composition is 1-15, preferably 3-10, such as about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15.

[0062] In certain embodiments, the weight portion of the Hovenia dulcis extract in the composition is about 20-50 parts, and the weight portion of the Nitraria tangutorum fructus extract is about 3-5 parts.

[0063] In certain embodiments, the weight portion of the Hovenia dulcis extract in the composition is about 50 parts, and the weight portion of the Nitraria tangutorum fructus extract is about 3 parts.

[0064] Application of the composition

[0065] The present invention provides the use of the "composition having a protective effect on blue light-induced melanocyte damage" as described above.

[0066] In certain embodiments, the composition is used to prepare food, cosmetics or medicines.

[0067] In certain embodiments, the composition is prepared into one or more selected from beverages, tablets, capsules, powders, candies or jelly.

[0068] In certain embodiments, the composition is used for preventing, inhibiting, reducing or eliminating skin pigmentation.

[0069] In certain embodiments, the composition is used for preventing, inhibiting, alleviating or eliminating skin wrinkles.

[0070] In certain embodiments, the composition is used to improve melasma.

[0071] In certain embodiments, the use is for non-therapeutic purposes.

[0072] In certain embodiments, the skin pigmentation, wrinkles, or melasma is caused by exposure to blue light.

[0073] In certain embodiments, the composition inhibits tyrosinase activity and / or reduces blue light-induced melanin production in skin cells.

[0074] Example

[0075] The present invention will be more easily understood with reference to the following experimental examples, which are included only to illustrate certain aspects and embodiments of the present invention and are not intended to limit the present invention.

[0076] Unless otherwise stated, the reagents used in this example are all commercially available materials or conventional materials.

[0077] Example 1 Preparation of Hovenia dulcis powder and Nitraria tangutorum fruit powder

[0078] Preparation of Hovenia dulcis powder:

[0079] A water extraction method is adopted, wherein the mass ratio of Hovenia dulcis to water in the water extraction step is 1:8, the heating temperature is 100°C, the extraction time is 2h, sieving is performed using a 400-mesh filter, vacuum concentration is performed using a vacuum degree of -0.08mpa, and then fine filtration is performed using a 100nm ceramic membrane. After high-temperature instantaneous sterilization, the extract is spray-dried, crushed, and sieved through an 80-mesh sieve to obtain a finished Hovenia dulcis extract.

[0080] Preparation of Nitraria tangutorum powder:

[0081] A water extraction method was adopted, wherein in the water extraction step, the mass ratio of white thorn fruit to water was 1:10, the heating temperature was 90°C, the extraction time was 2h, vacuum concentration was performed using a vacuum degree of -0.08mpa, and the white thorn fruit extract was obtained after spray drying and crushing and then sieved through an 80-mesh sieve.

[0082] Example 2 Detection of relative content of melanin and intracellular tyrosinase activity

[0083] instrument:

[0084] Incubator, ELISA reader, water bath, blue light source

[0085] Reagents:

[0086] 1×PBS solution, RPMI 1640 basic medium, RPMI 1640 complete medium (1640+10% serum+1% penicillin and streptomycin double antibodies), DMEM basic medium, DMEM complete medium (DMEM+10% serum+1% double antibodies), kojic acid, 1M NaOH (10% DMSO). B16 cells were cultured with DMEM medium for 1-2 generations before plating to allow the cells to produce melanin.

[0087] 2.1 Determination of melanin content:

[0088] ①Cultivate B16 cells in RPMI1640 for 24 h, switch to DMEM medium and culture B16 cells for at least one generation, then inoculate B16 cell suspension (1×10 5 cells / well) 1 mL (DMEM complete medium), cultured in a 37°C, 5% CO2 incubator for 24 h;

[0089] ② Aspirate the culture medium in the well plate, add 200μL PBS to wash once, add 200μL PBS to cover the cells in the plate, wrap the blank control well with tin foil, and irradiate the model group, positive control group and each treatment group with a blue light source with an illumination of 100Lux for 10min. After irradiation, aspirate the PBS in each well, add 1mL of DMEM complete culture medium to the 4 wells in the first column of the 24-well plate as a blank control, add 200μg / ml kojic acid to the second column as a positive control, and the rest of the columns are treatment groups;

[0090] ③ When the cell density reaches 60-70%, add the test substances with the same concentration but different ratios (DMEM complete medium is used as solvent). According to the CCK-8 test results of the three test substances, the total concentration with the lowest toxicity is selected as 31.25 μg / mL;

[0091] ④Discard the old culture medium and add 1 mL of PBS to the well plate for washing once;

[0092] ⑤ Add 500 μL of trypsin to each well and place in a 37°C incubator for 5 minutes to fully digest;

[0093] ⑥ Transfer the digested cells to a 1.5 ml centrifuge tube, wash once with 500 μl of PBS, transfer to a centrifuge tube, and 1×10 4 Centrifuge at rpm for 10 min;

[0094] ⑦Discard the supernatant, add 200 μL 1M NaOH (containing 10% DMSO), and place in a water bath at 80°C for about 30 minutes until the melanin is dissolved. Transfer 100 μL of the solution to a 96-well plate and read the plate at a wavelength of 405 nm or 475 nm.

[0095] Melanin relative content calculation formula:

[0096] Relative content of melanin = absorbance value of sample treatment group / absorbance value of blank control group.

[0097] 2.2 Intracellular tyrosinase activity detection:

[0098] ① Add PBS solution around the 96-well plate (26 wells);

[0099] ② Add 100 μL of DMEM complete medium to the middle 6 wells in the second column of the 96-well plate as a blank control.

[0100] ③ 100 μL of B16 cell suspension (10,000 cells / well) was inoculated into the middle 6 wells of columns 3-11 of a 96-well plate and cultured in a 37°C, 5% CO2 incubator for 24 h. The model group and sample group were modeled with blue light damage using the same method as above.

[0101] ④ Prepare 31.25 μg / mL of the test substance in different ratios (DMEM complete medium as solvent); use kojic acid as a positive control, and set different examples and comparative examples corresponding to the sample ratios (Table 1).

[0102] ⑤Discard the old culture medium, add 100 μL of the substance to be tested at a concentration of 31.25 μg / mL to columns 3-11 of the well plate, and culture for 24 to 48 hours;

[0103] ⑥Discard the old culture medium and add 100 μL PBS to columns 2-11 of the well plate to wash twice;

[0104] ⑦ Add 50 μL of 1% Triton X-100 solution to each well, quickly freeze in a -80°C refrigerator for 30 min, then thaw at room temperature to rupture the cells;

[0105] ⑧After warming at 37℃ for 5-10min, add 50μL of 10mM L-DOPA solution;

[0106] ⑨Incubate at 37℃ for 1-2h, and detect absorbance at 490nm. Calculate tyrosinase inhibition rate using the formula.

[0107] Tyrosinase inhibition rate = (1-average absorbance of sample group / average absorbance of model group) × 100%.

[0108] The test substances prepared using the Hovenia dulcis powder and Nitraria tangutorum fruit powder obtained in Example 1, the settings of the controls and the test results are shown in Table 1 below.

[0109] Table 1 Composition components and experimental results

[0110]

[0111] The results of the melanin content and intracellular tyrosinase activity are shown in Table 1 and Figure 1-2 It can be seen that when the specific ratio of Hovenia dulcis to Nitraria tangutorum is 5-50:3-10, the tyrosinase inhibition rate of the experimental example compared with the model group is 51.99%-73.53%; and when Hovenia dulcis powder is 50 parts and Nitraria tangutorum powder is 3 parts, Example 4 has the best tyrosinase inhibition effect compared with the model group, with an inhibition rate of 73.53% (Table 1 and Figure 1 ). When the specific ratio of Hovenia dulcis to Nitraria tangutorum was 5-50:3-10, the relative content of melanin in the embodiment group was 169.1%-105.4% compared with the model group; and when Hovenia dulcis 50 parts and Nitraria tangutorum 3 parts, the relative content of melanin production was the lowest at 105.4% (Table 1 and Figure 2 ).

Claims

1. A composition, characterized in that Includes Hovenia dulcis fruit extract and Nitraria tangutorum fruit extract.

2. The composition according to claim 1, comprising Hovenia dulcis fruit extract and Nitraria tangutorum fruit extract as active ingredients.

3. The composition according to claim 1 or 2, wherein the content of dihydromyricetin in the Hovenia dulcis extract is ≥ 0.5%.

4. The composition according to any one of claims 1 to 3, wherein the Hovenia dulcis extract is Hovenia dulcis powder; preferably, the Hovenia dulcis powder is a powder obtained by drying a water extract or an alcohol extract of Hovenia dulcis; preferably, the Hovenia dulcis powder is obtained by the following preparation method, which comprises the steps of heating water extraction or alcohol extraction, filtering, concentrating, filtering, drying, crushing, and optionally screening and mixing.

5. The composition according to any one of claims 1 to 4, wherein the Nitraria tangutorum fruit extract is Nitraria tangutorum fruit powder; preferably, the Nitraria tangutorum fruit powder is a powder obtained by drying a water extract or an alcohol extract of Nitraria tangutorum fruit; preferably, the Nitraria tangutorum fruit powder is obtained by the following preparation method, which comprises the steps of water extraction or alcohol extraction, concentration, drying, and optionally crushing and sieving.

6. The composition according to any one of claims 1-5, wherein the weight parts of the Hovenia dulcis extract are 5-50, and the weight parts of the Nitraria tangutorum fruit extract are 3-10; preferably, the weight parts of the Hovenia dulcis extract are 20-50, and the weight parts of the Nitraria tangutorum fruit extract are 3-5.

7. A product comprising the composition according to any one of claims 1 to 6; preferably, the product is selected from one or more of a beverage, a tablet, a soft capsule, a hard capsule, a powder, a granule, a candy or a jelly.

8. Use of the composition according to any one of claims 1 to 6 in the preparation of food, cosmetics or medicines; preferably, the food, cosmetics or medicines are in the form of one or more selected from the group consisting of drinks, tablets, soft capsules, hard capsules, powders, candies or jellies; preferably, the food, cosmetics or medicines are used to prevent, inhibit, reduce or eliminate skin pigmentation, or to improve chloasma.

9. Use of the composition according to any one of claims 1 to 6 or the product according to claim 7 for preventing, inhibiting, alleviating or eliminating skin pigmentation or for improving chloasma; preferably, the use is for non-therapeutic purposes; preferably, the skin pigmentation is induced by blue light.

10. The use according to claim 9, wherein the composition or product inhibits tyrosinase activity and reduces melanin generated by blue light.

11. A method for inhibiting tyrosinase activity or reducing blue light-induced melanin production, comprising the step of contacting the composition of any one of claims 1 to 6 or the product of claim 7 with cells; preferably, the cells are skin cells; preferably, the cells are melanocytes.

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