Composition for relieving blue light damage

Abstract

The invention belongs to the technical field of functional nutrient substance research, and particularly relates to a composition for relieving blue light damage. The composition is prepared from the following necessary components: 1, 3-dioleic acid-2-palmitin and taurine, and in the composition, the mass ratio of the 1, 3-dioleic acid-2-palmitin to the taurine is 500: (0.38 to 13.42). Compared with single OPO or taurine, the composition provided by the invention can more effectively relieve blue light damage; particularly, active oxygen free radicals generated by blue light irradiation can be effectively eliminated, the content of lipid oxidation products of eyes can be reduced, the activity of antioxidant enzymes can be increased, the relative expression quantity of proinflammatory factor IL-1beta gene il-1beta can be reduced, the occurrence of inflammation can be inhibited, and the structure and state of damaged tissues of the eyes can be improved.

Description

[0001] This application is a divisional application of the Chinese invention patent application with application number 202410903854.4, application date July 8, 2024, and invention name “A composition for alleviating blue light damage”. Technical Field

[0002] The invention belongs to the technical field of functional nutrient research, and specifically relates to a composition for alleviating blue light damage. Background Art

[0003] Blue light refers to short-wavelength light with a wavelength between 400nm and 500nm. It is an important component of natural light. Blue light plays an important role in maintaining normal vision and human health, and plays an important role in visual perception and color vision. Blue light is also involved in pupil light reflex and pupil contraction, and is also involved in regulating the biological clock. Therefore, appropriate blue light exposure is beneficial to health, such as improving alertness, improving memory function, regulating emotions and increasing happiness. But on the other hand, blue light has high energy and strong penetrating power, so it can penetrate the lens and reach the retina, causing blue light damage to the retina. Blue light damage is mainly manifested in accelerating oxidative damage to cells in the macular area of ​​the retina, causing retinal degenerative changes, forming photoretinitis, causing irreversible damage to the retina of the human eye, and even blindness. In addition to blue light from natural sources, artificial light sources such as light emitting diodes (LEDs) are also an important source of blue light. LEDs mainly excite yellow light phosphors through blue light chips to emit white light. Therefore, under high color temperature conditions, there will be a strong blue light peak in the light source spectrum. With the rapid development of related technologies, LED currently has the advantages of long life, small size, high brightness, and low energy consumption. Because of this, it is widely used in daily life, including children's toys, various lamps and various screen electronic products. According to statistics, 90% of optoelectronic toys on the market currently use LED; and whether it is the need for online learning or the need for regular teaching and learning, in many cases there will be teaching or learning links through desktop computers, tablets or even smart phones, so children will be exposed to a large amount of blue light in the process of participating in such teaching activities. However, children have insufficient cognitive ability about the hazards of light sources, and due to their natural curiosity, they are more susceptible to blue light damage, causing irreversible damage to their eyesight.

[0004] The protection methods for blue light mainly include physical protection and chemical protection methods. Physical protection mainly uses anti-blue light glasses, anti-blue light screens, anti-blue light films, etc. to physically block blue light. Physical means can indeed block some blue light from reaching the human eye, which can alleviate the symptoms of eye fatigue when using digital devices to a certain extent, and may also protect the retina from the damage of blue light, but at the same time it also interferes with the positive effects of blue light on normal visual function and physiological regulation. Chemical protection methods mainly involve taking antioxidants, free radical scavengers and even anti-inflammatory drugs for related protection and treatment. Since retinal cells and lens epithelial cells produce free radicals such as singlet oxygen or superoxide anions after being irradiated by blue light, oxidative stress is exerted on eye cells and tissues. Antioxidants or free radical scavengers can inhibit the accumulation of oxidative stress effects. For example, lutein can protect cells from photooxidation and photodamage, and can be used as an active high-energy blue light filter to provide antioxidant protection for organisms. Vitamin E can remove reactive oxygen free radicals (ROS) generated by blue light exposure to a certain extent, while vitamin C can also prevent cellular oxidative damage to a certain extent, thereby preventing blue light exposure from damaging mitochondrial DNA of cells. While blue light exposure causes the generation and accumulation of ROS, it can also promote the release of inflammatory factors, which suggests that the use of anti-inflammatory drugs can also alleviate the damage to the eyes caused by blue light exposure. Chemical protection methods will not interfere with the normal physiological function of blue light, but can also alleviate the damage caused by blue light in a targeted manner.

[0005] 1,3-dioleyl-2-palmitoylglycerol (OPO) is an important component of breast milk lipids, and its palmitic acid is located at the Sn-2 position. In the digestive tract, the fatty acids at the Sn-1 and Sn-3 positions are released under the decomposition of pancreatic lipase to form free fatty acids, while the palmitic acid at the Sn-2 position is still attached to the glycerol group and absorbed by the intestine. Therefore, no free palmitic acid is produced in this process, and it will not form insoluble soaps with cations in the intestine, such as calcium ions and magnesium ions, which will affect the baby's absorption of calcium, etc., and therefore will not make the baby's stool hard, increasing the risk of constipation and abdominal pain. Because of this, OPO structured fats are currently widely added to infant formula milk powder to help improve the absorption rate of nutrients in milk powder, reduce discomfort such as constipation, and further simulate the function of breast milk. At present, no public reports have been found on OPO in repairing blue light damage.

[0006] Taurine is a stable endogenous sulfur-containing amino acid present in organisms. Taurine is widely distributed in various tissues and organs of humans and animals, with higher content in skeletal muscle and heart. In addition, a certain amount of taurine is also present in the milk of mammals, which is an essential nutrient for the growth and development of infants and young children. Studies have shown that taurine, as a conditionally essential amino acid for the human body, has multiple biological functions such as enhancing immunity, relieving fatigue, anti-oxidation, promoting fat metabolism, protecting the nervous system, and improving memory. Reference 1 discloses a health food for relieving visual fatigue and a preparation method thereof, wherein the health food composition contains taurine, zinc gluconate, lutein, bilberry extract, vitamin E, and vitamin C, etc. However, it has not confirmed the actual effect of the relevant combination in relieving visual fatigue through experiments, nor has it proved whether there is a synergistic effect between the components. Reference 2 discloses a composition of an anti-blue light eye spray, which contains 0.05-0.1% orange peel dry powder, 0.01-0.03% lutein powder, 0.05-0.1% vitamin A, 0.1-0.2% tea polyphenol powder, 0.05-0.1% grape seed extract powder, 0.05-0.2% taurine, etc. However, it does not verify and publish the specific effect of the above composition in resisting blue light damage to the eyes, nor does it confirm whether there is a synergistic effect between the components.

[0007] References:

[0008] Reference 1: CN108771243A;

[0009] Reference 2: CN109549947A. Summary of the invention

[0010] Problem that the invention aims to solve

[0011] Although there have been studies in the prior art on compositions that are beneficial to the eyes, such as relieving visual fatigue or resisting blue light, such research still cannot be said to be perfect, and there is still room for development of chemical protective measures to alleviate blue light damage.

[0012] The purpose of the present invention is to provide a composition for alleviating blue light damage using OPO and taurine. By adopting a zebrafish blue light damage model, the present invention verifies the effects of OPO and taurine respectively and their combination in alleviating blue light damage, and unexpectedly finds that when the two are combined in a certain proportion, an effect that cannot be achieved when used alone can be obtained, that is, the present invention finds that OPO and taurine have a synergistic effect in alleviating blue light damage.

[0013] Solutions for solving problems

[0014] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0015] The first aspect of the present invention provides a composition, wherein the composition comprises the following essential components:

[0016] 1,3-dioleyl-2-palmitoylglycerol and taurine;

[0017] Furthermore, in the composition, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol to the taurine is 500:(0.38-42.25).

[0018] In some embodiments, in the composition, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol to the taurine is 500:(13.42-42.2).

[0019] The second aspect of the present invention provides use of the composition described in the first aspect of the present invention in preparing food that helps alleviate blue light damage.

[0020] In some embodiments, the method of helping to alleviate blue light damage includes at least one of reducing ocular cell apoptosis, reducing the accumulation of ocular reactive oxygen free radicals, reducing the content of ocular lipid oxidation products, increasing the activity of ocular antioxidant enzymes, reducing the expression of gene il-1β, and improving ocular tissue structure.

[0021] In some embodiments, the lipid oxidation product comprises malondialdehyde.

[0022] In some embodiments, the antioxidant enzyme comprises catalase.

[0023] In some embodiments, the improving eye tissue structure includes at least one of restoring eye size, arranging the retina in an orderly hierarchical structure, thickening the pigment epithelium without pigment diffusion, arranging the cone and rod cell layers in a regular manner, and arranging the outer nuclear layer and inner nuclear layer cells in a regular manner.

[0024] In some embodiments, the food is a solid dairy product or a liquid dairy product.

[0025] In some embodiments, the food is infant food or children's food.

[0026] In some embodiments, the food further contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients, and any acceptable excipients.

[0027] Effects of the Invention

[0028] Through the implementation of the above technical solution, the present invention achieves the following technical effects:

[0029] The present invention has developed a composition through extensive research, which contains OPO and taurine in a specific ratio. Compared with OPO or taurine alone, the composition provided by the present invention can more effectively alleviate blue light damage; in particular, it can effectively remove reactive oxygen free radicals generated by blue light irradiation, reduce the content of lipid oxidation products such as MDA in the eye, increase the activity of antioxidant enzymes such as CAT, reduce the relative expression of the pro-inflammatory factor IL-1β gene il-1β, inhibit the occurrence of inflammation, and improve the structure and state of damaged eye tissues. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 : Fluorescence intensity of apoptotic cells in zebrafish eyes after sample treatment in Example 1; compared with the model control group, **p<0.01, ***p<0.001.

[0031] Figure 2 : A typical graph of the fluorescence intensity of apoptotic cells in the eyes of zebrafish after sample treatment in Example 1; wherein the fluorescent particles indicated by the arrows are apoptotic cells.

[0032] Figure 3 : Fluorescence intensity of apoptotic cells in zebrafish eyes after sample treatment in Example 1; compared with the model control group, **p<0.01, ***p<0.001; compared with OPO 250μg / mL, # p<0.05; compared with taurine 0.403μg / mL, □ p<0.05; compared with taurine 1.34μg / mL, & p<0.05; compared with taurine 6.71μg / mL, ▲ p<0.05; compared with taurine 20.1μg / mL, ○ p<0.05.

[0033] Figure 4 : A typical graph of the fluorescence intensity of apoptotic cells in the eyes of zebrafish after sample treatment in Example 1; wherein the fluorescent particles indicated by the arrows are apoptotic cells.

[0034] Figure 5 : Average fluorescence intensity of zebrafish head after sample treatment in Example 2; Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001; Compared with OPO 250μg / mL concentration, # p<0.05; compared with taurine 20.1μg / mL concentration, ∞ p<0.01.

[0035] Figure 6 : A typical graph of the average fluorescence intensity of the zebrafish head after sample treatment in Example 2; wherein the dotted box is the analysis area of ​​the zebrafish head.

[0036] Figure 7 : The MDA content of zebrafish after sample treatment in Example 3; Compared with the model control group, **p<0.01, ***p<0.001; Compared with taurine 20.1μg / mL concentration, ○ p<0.05.

[0037] Figure 8 : The CAT activity of zebrafish after sample treatment in Example 3; Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001; Compared with taurine 20.1 μg / mL concentration, ○ p<0.05.

[0038] Fig. 9 : Relative expression of zebrafish il-1β gene after sample treatment in Example 4; wherein, compared with the model control group, *p<0.05.

[0039] Fig.10 : Relative expression of zebrafish tnf-α gene after sample treatment in Example 4; compared with the model control group, ***p<0.001.

[0040] Fig.11 : Example diagram of zebrafish eye tissue analysis area and eye retinal cross-sectional slices in Example 5; wherein, the black single-sided square bracket is the zebrafish eye retina.

[0041] Fig.12 : Typical diagram of zebrafish eye tissue structure after sample treatment in Example 5; wherein ① is a white cross double arrow line, indicating the sum of the dimensions of the anterior-posterior axis (A / P) and the dorsal-ventral axis (D / V) of the eye; ② is a black half square bracket, indicating the retinal structure; ③ is a black arrow, indicating the pigment epithelium; ④ is a white dotted elliptical area, indicating the cone and rod cell layers; ⑤ is a black dotted fan-shaped area, indicating the outer nuclear layer and the inner nuclear layer. DETAILED DESCRIPTION

[0042] The embodiments of the present invention are described below, but the present invention is not limited thereto. The present invention is not limited to the various structures described below, and various changes can be made within the scope of the present invention, and the embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention.

[0043] I. Definition of Terms

[0044] In the present invention, a numerical range expressed using “a value A to a value B”, “a value A - a value B”, “above a value A” or below a value A means a range including the endpoints A and B.

[0045] In the present invention, the term "comprising", "including" or "containing" may mean inclusive or open-ended, and does not exclude additional, uncited components or method steps. At the same time, "comprising", "including" or "containing" may also mean closed, excluding additional, uncited components or method steps.

[0046] In the present invention, the term "about" is used to define the numerical range and parameters of the present invention are approximate values, and the specific relevant values ​​are presented as accurately as possible. Unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in the present invention are modified by "about". Here, "about" usually means that the actual value is within ±3%, ±2%, ±1% or ±0.5% of a particular value or range.

[0047] In the present invention, the term "normal temperature" or "room temperature" means an indoor ambient temperature of "23±2°C".

[0048] In the present invention, "infant" is used to refer to a human group of 0 to 6 months old.

[0049] In the present invention, "older infants" are used to refer to a human group of 6 to 12 months of age.

[0050] In the present invention, the term "infant" is used to refer to a human group of 12 to 36 months old.

[0051] In the present invention, "infants" are used to refer to a human group under the age of 3.

[0052] In the present invention, the term "children" used herein refers to a human living group that is older than 3 years old and younger than 12 years old and is in the growth and development stage.

[0053] In the present invention, the "infant formula" includes infant formula, follow-up infant formula and toddler formula. Generally, infant formula is used as a breast milk substitute from birth, follow-up infant formula is used as a breast milk substitute from 6 to 12 months after the birth of the infant, and toddler formula refers to a breast milk substitute from 12 to 36 months after the birth of the infant.

[0054] In the present invention, "animal milk" is used to refer to the liquid obtained from the mammary gland of a mammal in lactation. The term "animal milk" should be interpreted broadly and covers both raw milk (i.e., liquid obtained directly from the mammary gland) and standardized milk products (such as, for example, skim milk or whole milk).

[0055] Unless otherwise defined, other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0056] II. Composition

[0057] The present invention provides a composition comprising the following essential components: 1,3-dioleyl-2-palmityl glycerol and taurine; and in the composition, the mass ratio of the 1,3-dioleyl-2-palmityl glycerol to the taurine is 500:(0.38-42.25).

[0058] Compared with 1,3-dioleyl-2-palmitoylglycerol or taurine alone, the composition provided by the present invention can more effectively alleviate the damage caused by blue light to the eyes. In terms of alleviating blue light damage, 1,3-dioleyl-2-palmitoylglycerol and taurine have a synergistic effect, especially when the content ratio of the two is within the above range, the relevant effect is particularly prominent.

[0059] Exemplarily, in the composition, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol to the taurine can be about 500:0.4, 500:0.5, 500:1, 500:2, 500:3, 500:4, 500:5, 500:6, 500:7, 500:8, 500:9, 500:10, 500:11, 500:12, 500:13, 500:14, 500:15, 500:16, 500:17, 500:18, 500:19, 500:20, 500:21, 500:22, 500:23, 500:24, 500:25 0:18, 500:19, 500:20, 500:21, 500:22, 500:23, 500:24, 500:25, 500:26, 500:27, 500:28, 500:29, 500:30, 500:31, 500:32, 500:33, 500:34, 500:35, 500:36, 500:37, 500:38, 500:39, 500:40, 500:41 or 500:42, etc.

[0060] In order to obtain a better synergistic effect in alleviating blue light damage, in some embodiments, in the composition, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol ester to the taurine is 500:(0.403-42.2); preferably 500:(6.71-42.2); more preferably 500:(13.42-42.2); even more preferably 500:(20.1-42.2); further more preferably 500:(38.2-42.2).

[0061] In some embodiments, the composition consists of 1,3-dioleyl-2-palmitoylglycerol and taurine, and in the composition, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol to the taurine is 500:(0.38-42.25).

[0062] The present invention does not particularly limit the form of the composition, and typically, the composition may be liquid or solid.

[0063] In some embodiments, in the composition described in the present invention, the content of 1,3-dioleoyl-2-palmitoylglycerol is greater than or equal to 200 μg / mL, preferably greater than or equal to 250 μg / mL, and more preferably 250-500 μg / mL; the content of taurine is greater than or equal to 0.1 μg / mL, preferably greater than or equal to 0.403 μg / mL, more preferably greater than or equal to 6.71 μg / mL, and even more preferably 6.71-20.1 μg / mL.

[0064] The composition of the present invention is usually an artificially synthesized or compounded composition, that is, it is not a natural composition such as breast milk.

[0065] III. Uses to alleviate blue light damage

[0066] The present invention proposes to compound 1,3-dioleyl-2-palmitoylglycerol and taurine in a specific ratio to alleviate the damage of blue light to the eyes, and the two substances have a synergistic effect. Therefore, the present invention provides the use of the above-mentioned composition in the preparation of food that helps to alleviate blue light damage, and the blue light damage alleviation described in the present invention is not for the purpose of treating or preventing diseases, and the blue light damage does not reach the level of disease.

[0067] In some embodiments, the method of helping to alleviate blue light damage includes at least one of reducing ocular cell apoptosis, reducing the accumulation of ocular reactive oxygen free radicals, reducing the content of ocular lipid oxidation products, increasing the activity of ocular antioxidant enzymes, reducing the expression of gene il-1β, and improving ocular tissue structure.

[0068] The present invention has found through experiments that the composition of 1,3-dioleyl-2-palmitoylglycerol and taurine can effectively remove the reactive oxygen free radicals generated by blue light irradiation in the eyes of zebrafish, reduce the content of lipid oxidation products in the eyes, increase the activity of antioxidant enzymes, reduce the relative expression of the pro-inflammatory factor IL-1β gene il-1β, inhibit the occurrence of inflammation, and improve the structure and state of damaged eye tissues. The two components in the composition have a synergistic effect in the above effects.

[0069] In some embodiments, the lipid oxidation product comprises malondialdehyde (MDA).

[0070] In some embodiments, the antioxidant enzyme comprises catalase (CAT).

[0071] In some embodiments, the improving eye tissue structure includes at least one of restoring eye size, arranging the retina in an orderly hierarchical structure, thickening the pigment epithelium without pigment diffusion, arranging the cone and rod cell layers in a regular manner, and arranging the outer nuclear layer and inner nuclear layer cells in a regular manner.

[0072] The present invention does not specifically limit the food with the effect of alleviating blue light damage that can be prepared using the above composition. In some embodiments, the food is a solid dairy product or a liquid dairy product; for example, milk powder, cheese, yogurt, liquid milk, etc. In some embodiments, the food is an infant food or a children's food; for example, infant formula milk powder, children's formula milk powder, hydrolyzed formula milk powder, children's snacks, etc.

[0073] In some embodiments, in the food, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol ester to the taurine is 500:(0.38-42.25); preferably 500:(0.403-42.2); more preferably 500:(6.71-42.2); further preferably 500:(13.42-42.2); even more preferably 500:(20.1-42.2); further more preferably 500:(38.2-42.2).

[0074] In food, the specific content of 1,3-dioleyl-2-palmitoylglycerol and taurine is not particularly limited, as long as it meets the requirements of food-related laws and regulations. In some embodiments, in the food, the content of 1,3-dioleyl-2-palmitoylglycerol is greater than or equal to 3500 μg / mL, preferably greater than or equal to 5500 μg / mL, and more preferably 5500-7800 μg / mL; the content of taurine is greater than or equal to 25 μg / mL, preferably greater than or equal to 35 μg / mL, more preferably greater than or equal to 49 μg / mL, and even more preferably 49-60 μg / mL.

[0075] Depending on the type of food and the final needs of the applicable object, in some embodiments, the food also contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients and any acceptable excipients.

[0076] As for the plant product ingredients, examples include fruits such as figs, pomegranates, kiwis, tangerines, oranges, pineapples, strawberries, apples, bananas, grapes, pears, cherries, blueberries, blackberries, blackcurrants, cranberries, raspberries, melons, emblica chinensis and bilberry or their extracts; fruit and vegetable substances such as onions, cucumbers, tomatoes, cauliflower, carrots, spinach, kale, Brussels sprouts, garlic, basil, oregano or their extracts; rice (indica rice , japonica rice, glutinous rice), wheat (wheat, barley, oats, rye), corn, sorghum, millet, sorghum, yellow rice, buckwheat, soybean, broad bean, pea, mung bean, red bean, kidney bean and other cereals or their extracts; walnut, pistachio, cashew, hazelnut, almond, apricot, pine nut, peanut, melon seed, chestnut, macadamia, ginkgo and other nuts or their extracts; coffee or its extract; and some edible and medicinal plant-based Chinese medicinal materials or their extracts.

[0077] As for animal dairy ingredients, examples include fresh milk from cows, sheep, etc., as well as reprocessed dairy products such as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, and hydrolyzed whey protein powder.

[0078] Examples of animal meat ingredients include meat ingredients from pigs, cattle, sheep, aquatic animals or poultry.

[0079] Functional added ingredients include vitamin supplements, mineral supplements, nucleotide supplements, dietary fiber, etc.

[0080] As for any acceptable excipients, examples include solvents, antioxidants, antibacterial agents, thickeners, diluents, cosolvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweeteners, edible flavors, edible colorings, etc.

[0081] Example

[0082] The embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out under normal conditions or conditions recommended by the manufacturer. Unless otherwise specified, the materials or instruments used are all commercially available conventional products.

[0083] Example 1: Evaluation of anti-apoptosis efficacy

[0084] 1. Test materials

[0085] 1.1. Sample preparation information

[0086] 1,3-dioleoyl-2-palmitoylglycerol (OPO) was purchased from Bangilodex, and the solvent was DMSO.

[0087] Taurine was purchased from DSM, and the solvent was standard dilution water.

[0088] Positive control: Lutein, purchased from DSM, solvent was DMSO.

[0089] 1.2. Experimental animals

[0090] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), and were bred and provided by Hangzhou Huante Biological Fish Farming Center. The experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0091] When conducting the experiment, the method of administering OPO monomer to zebrafish is: first dissolving OPO in DMSO as a mother solution, then diluting the mother solution with standard dilution water to the target concentration by utilizing the miscibility of DMSO and water, and raising the zebrafish in the sample solution of the target concentration; the method of administering taurine monomer to zebrafish is: dissolving taurine to the target concentration by using standard dilution water, and raising the zebrafish in the sample solution of the target concentration; the method of administering lutein monomer to zebrafish is the same as the method of administering OPO monomer to zebrafish; the method of administering OPO and taurine combination to zebrafish is: first dissolving OPO in DMSO as a mother solution, then utilizing the miscibility of DMSO and water, diluting the OPO mother solution with standard dilution water while dissolving taurine until the target concentration, and raising the zebrafish in the sample solution of the target concentration.

[0092] 1.3. Instruments, consumables and reagents

[0093] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China); electric focus continuous zoom fluorescence microscope (AZ100, Nikon, Japan); ultrasonic cleaning machine (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); fully automatic sample rapid grinding machine (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); blue light instrument (50w450nm, China).

[0094] Dimethyl sulfoxide (DMSO, Sigma, Switzerland; I2229063, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Pronase E (Shanghai Yuanye Biotechnology Co., Ltd., China); Acridine orange (AO, Shanghai MacLean Biochemical Technology Co., Ltd., China); Methylcellulose (Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0095] 2. Test methods

[0096] 2.1. Determination of maximum detectable concentration (MTC)

[0097] Wild-type AB zebrafish 1 day after fertilization (1dpf) were randomly selected and exposed to blue light after membrane rupture to establish a zebrafish blue light eye damage model. Model zebrafish with good development at 3dpf were selected and randomly distributed in 6-well plates, and 30 zebrafish were treated in each well (experimental group). Samples were dissolved in water (concentrations shown in Table 1-1), and normal control group and model control group were set up at the same time, with a capacity of 3mL per well. After 1 day of treatment at 28℃, the MTC of the samples on the model zebrafish was measured.

[0098] 2.2. Evaluation of anti-apoptosis efficacy

[0099] 2.2.1 Evaluation of the anti-apoptotic efficacy of monomer components

[0100] Randomly select 1dpf wild-type AB strain zebrafish and irradiate with blue light after membrane rupture to establish the zebrafish blue light eye damage model. At 3dpf, select model zebrafish with good development status and randomly distribute them in 6-well plates, and treat 30 zebrafish in each well (experimental group). Different concentrations of samples (see Table 1-2) were given in water: 125μg / ml, 250μg / ml, 500μg / ml, 1000μg / ml and 2000μg / ml, and the positive control lutein concentration was set to 1000μg / mL. At the same time, a normal control group and a model control group were set, and the volume of each well was 3mL. After 1 day of treatment at 28℃, the zebrafish in each experimental group were stained with AO light-proof for 30min, washed with standard dilution water 3 times, and 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. Image J software was used to analyze and collect data, and the fluorescence intensity of apoptotic cells in the eyes of zebrafish was analyzed and statistically analyzed. The anti-apoptotic effect of the samples was evaluated by the statistical analysis results of the above indicators. The statistical results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0101] 2.2.2 Evaluation of the anti-apoptotic efficacy of the composition

[0102] According to the above anti-apoptosis efficacy evaluation results, the concentrations of OPO and taurine were preferably used for combined experiments to evaluate the relevant effects of the composition. The experimental conditions were the same as the above evaluation method, and the positive control lutein concentration was set to 1000 μg / mL. A normal control group and a model control group were also set. After the experiment, the zebrafish treatment, data collection and analysis methods were the same as above.

[0103] 3. Test results

[0104] 3.1.MTC

[0105] See Table 1-1 for details. The test results show that the MTC of the positive control lutein, OPO and taurine for the model zebrafish is 2000μg / mL.

[0106] Table 1-1. Results of the concentration exploration experiment on the anti-apoptotic effect of samples (n=30)

[0107]

[0108] 3.2. Evaluation of anti-apoptosis efficacy

[0109] 3.2.1 Evaluation of the anti-apoptotic efficacy of monomer components

[0110] The experimental results showed that the fluorescence intensity of apoptotic cells in the eyes of 1dpf wild-type AB strain zebrafish increased significantly after the membrane was broken and irradiated with blue light (Table 1-2, Figure 1 ), and compared with the normal control group, p < 0.001, which indicates that blue light irradiation leads to a significant increase in apoptosis of zebrafish eye cells, that is, the modeling is successful. The positive control lutein, monomer component OPO and taurine all showed anti-apoptotic effects at certain concentrations (compared with the fluorescence intensity of apoptotic cells in the eye of the model control group, p < 0.01 or 0.001), the specific concentrations are lutein 1000μg / mL, OPO 500μg / mL, 1000μg / mL and 2000μg / mL and taurine 1000μg / mL and 2000μg / mL (see Table 1-2 for details). From the typical graph of the fluorescence intensity of apoptotic cells in the zebrafish eye ( Figure 2 ) It can also be found that the fluorescence intensity / spot size of the apoptotic cells in the eyes treated with the above-mentioned components at a specific concentration tends to weaken / become smaller compared with the model control group, and the higher the concentration of the monomer component, the more obvious the above effect.

[0111] Table 1-2. Results of the evaluation experiment on the anti-apoptotic effect of monomer components (n=10)

[0112]

[0113] 3.2.2 Evaluation of the anti-apoptotic efficacy of the composition

[0114] According to the evaluation results of the anti-apoptotic efficacy of the above monomer components, we selected different concentrations of OPO and taurine for combination (specific combination concentrations are shown in Tables 1-3), and evaluated their anti-apoptotic efficacy again. The results are shown in Tables 1-3 and Figure 3 . Under the conditions of this experiment, we found that the OPO monomer concentration of 500μg / mL can still effectively reduce the fluorescence intensity of zebrafish eye cells. However, since the selected concentration of taurine is much lower than the effective concentration in the monomer test (1000μg / mL), it cannot significantly reduce the fluorescence intensity of apoptotic cells in the eye, and therefore does not have the effect of preventing blue light damage. The combined OPO and taurine (the ratio of the two is 500: (0.403~40.2)) both showed significant anti-apoptotic effects on zebrafish eyes (compared with the model control group, p < 0.01 or 0.001). Moreover, the fluorescence intensity of apoptotic cells in the zebrafish eyes under the action of the combination is lower than the average fluorescence intensity of the cells when the two are used alone at the same concentration. OPO and taurine have a synergistic effect. When the ratio of OPO and taurine is 500: (0.403~40.2), the fluorescence intensity of apoptotic cells in the zebrafish eyes is reduced to a very low level (such as Figure 3 The size and / or intensity of the fluorescent spots shown in the typical fluorescence intensity graph all tend to become smaller and / or weaker (as shown in Figure 2). Figure 4 As shown), this indicates that the above combination has a good effect of resisting blue light-induced ocular cell apoptosis. In particular, when the combined concentration of the two is 250+20.1 μg / mL, the composition has a very obvious effect of resisting ocular cell apoptosis, which is better than the effect of OPO concentration of 250 μg / mL and taurine concentration of 20.1 μg / mL when used alone (p < 0.05), and the effect of the combination is significantly better than the average effect of the two when used alone (p < 0.05). It can be considered that the synergistic effect of the two at this ratio (500:40.2) is the most significant.

[0115] Table 1-3. Results of the experimental evaluation of the anti-apoptotic efficacy of the composition (n=10)

[0116]

[0117] Example 2: Evaluation of ROS scavenging effect

[0118] 1. Test materials

[0119] 1.1. Sample preparation information

[0120] The solvent of OPO is DMSO, the solvent of taurine is standard dilution water, and the solvent of the positive control lutein is also DMSO.

[0121] 1.2. Experimental animals

[0122] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), and were bred and provided by Hangzhou Huante Biological Fish Farming Center. The experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0123] When conducting the experiment, the method of administering different samples to zebrafish was the same as described in Example 1.

[0124] 1.3. Instruments, consumables and reagents

[0125] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China); electric focus continuous zoom fluorescence microscope (AZ100, Nikon, Japan); ultrasonic cleaning machine (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); blue light instrument (50w450nm, China).

[0126] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Pronase E (Shanghai Yuanye Biotechnology Co., Ltd., China); Cell ROX Green Reagent (Invitrogen, USA); Methylcellulose (Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0127] 2. Detection Method

[0128] The zebrafish of the Albino strain with melanin allele mutant at 1dpf were randomly selected and irradiated with blue light to establish the zebrafish blue light eye damage model. At 3dpf, the model zebrafish with good development status were selected and randomly distributed in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples and positive control lutein (concentrations are shown in Table 2-1) were given water-soluble respectively, and the normal control group and model control group were set up at the same time, with a capacity of 3mL per well. After being treated at 28℃ for 1 day, the zebrafish in each experimental group were stained with CellROX in the dark for 30min, washed with standard dilution water 3 times, and 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software, and the average fluorescence intensity of the zebrafish head was analyzed and statistically analyzed. The statistical analysis results of this indicator were used to evaluate the effect of the sample on the ROS level. The statistical analysis results are expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0129] 3. Test results

[0130] Blue light irradiation leads to the accumulation of reactive oxygen free radicals (ROS) in eye cells, causing oxidative damage to cells. The test results showed that after blue light irradiation, the fluorescence intensity of ROS in the model control group was significantly higher than that in the normal control group (p < 0.001), indicating that blue light irradiation caused abnormal accumulation of ROS in the zebrafish eye, and the modeling was successful. Compared with the model control group, the positive control lutein (1000μg / mL), OPO (250μg / mL), taurine (20.1μg / mL) and OPO+taurine combination (250+20.1μg / mL) all had ROS scavenging effects, which were specifically manifested in reducing the average fluorescence intensity of the zebrafish head (Table 2-1, Figure 5 ), and the fluorescence level of the zebrafish head was also lower than that of the model control group ( Figure 6); Among them, the positive control lutein (1000μg / mL), OPO (250μg / mL) and OPO+taurine combination (250+20.1μg / mL) have a significant level of scavenging effect on ROS compared with the model control group (p<0.05 or p<0.01). Compared with the monomer components OPO and taurine, the combination (OPO 250μg / mL+taurine 20.1μg / mL, that is, the ratio of the two is 500:40.2) has a better effect in scavenging ROS generated by blue light irradiation (compared with OPO 250μg / mL concentration, p<0.05; compared with taurine 20.1μg / mL concentration, p<0.01), and the effect of the combination is significantly better than the average value of the effects of the two monomers at the same concentration of the combination (p<0.01). OPO and taurine have a synergistic effect when used in this ratio, which can enhance the effect of the monomer in scavenging ROS generated by blue light irradiation.

[0131] Table 2-1. Sample ROS scavenging effect evaluation test results (n = 10)

[0132]

[0133] Example 3: Evaluation of the effect on the content / activity of enzymes related to oxidative damage

[0134] 1. Test materials

[0135] 1.1 Sample configuration information

[0136] The solvent of OPO was DMSO, and the solvent of taurine was standard dilution water.

[0137] 1.2 Experimental animals

[0138] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), and were bred and provided by Hangzhou Huante Biological Fish Farming Center. The experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0139] When conducting the experiment, the method of administering different samples to zebrafish was the same as described in Example 1.

[0140] 1.3 Instruments, consumables and reagents

[0141] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China); ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); blue light instrument (50w450nm, China); 96-well plate (Nest Biotech, China); fully automatic sample rapid grinding instrument (JXFSTPRP-24L, Shanghai Jingxin Laboratory Equipment Technology Department, China); high-speed refrigerated centrifuge (Heraeus Fresco17, ThermoFisher, Germany); multi-function microplate reader (SPARK, TECAN, Austria); electronic constant temperature stainless steel water bath (HHS-2S, Shanghai Kanglu Instrument Equipment Co., Ltd., China).

[0142] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); pronase E (Shanghai Yuanye Biotechnology Co., Ltd., China); BCA protein concentration assay kit (Boster Biotechnology, China); sodium chloride injection (Hunan Kelun Pharmaceutical Co., Ltd., China); PBS phosphate buffer (Biosharp, China); glacial acetic acid (Shanghai Aladdin Biochemical Technology Co., Ltd., China); malondialdehyde (MDA) kit (Nanjing Jiancheng Bioengineering Institute, China); hydrogen peroxide (CAT) assay kit (Nanjing Jiancheng Bioengineering Institute, China).

[0143] 2. Detection Method

[0144] 2.1 Effect on malondialdehyde (MDA) content

[0145] Randomly select 1dpf wild-type AB strain zebrafish and irradiate with blue light after membrane rupture to establish a zebrafish blue light eye damage model. At 3dpf, model zebrafish with good development status were selected and randomly distributed in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples were dissolved in water (concentrations are shown in Table 3-1), and a normal control group and a model control group were set up at the same time. The volume of each well was 3mL, and 3 biological replicates were set up in parallel. After 1 day of treatment at 28℃, zebrafish samples were collected according to the instructions of the MDA kit, and the MDA content was detected using a multifunctional microplate reader. The MDA content in zebrafish was analyzed, and the statistical analysis results of this indicator were used to evaluate the effect of the sample on the MDA content. The statistical analysis results are expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0146] 2.2 Effect on catalase (CAT) activity

[0147] Randomly select 1dpf wild-type AB strain zebrafish and irradiate with blue light after membrane rupture to establish a zebrafish blue light eye damage model. At 3dpf, model zebrafish with good development status were selected and randomly distributed in 6-well plates, and 30 zebrafish were treated in each well (experimental group). The samples were dissolved in water (concentrations are shown in Table 3-2), and a normal control group and a model control group were set up at the same time. The volume of each well was 3mL, and 3 biological replicates were set up in parallel. After 1 day of treatment at 28℃, zebrafish samples were collected according to the instructions of the CAT kit, and CAT activity was detected using a multifunctional microplate reader to analyze the CAT activity in zebrafish. The statistical analysis results of this indicator were used to evaluate the effect of the sample on CAT activity. The statistical analysis results are expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0148] 3. Test results

[0149] 3.1 Effect on MDA content

[0150] Blue light irradiation causes oxidative damage to the eye, including oxidation of cellular lipids, and malondialdehyde (MDA) is one of the lipid oxidation products, which can measure the severity of oxidative damage. The experimental results show that after blue light irradiation, the MDA content in the model control group increased significantly compared with normal cells (p<0.001), indicating that blue light irradiation caused obvious oxidative damage to eye cell lipids, and lipid oxidation caused a significant increase in MDA content. Compared with the model control group, monomeric OPO (250μg / mL), monomeric taurine (20.1μg / mL) and the combination of OPO and taurine (250+20.1μg / mL) all reduced the MDA content in the eyes of the model zebrafish (as shown in Table 3-1, Figure 7 As shown in the figure, the effects of monomeric OPO (250 μg / mL) and the combination of OPO and taurine (250+20.1 μg / mL) on reducing MDA were more significant (compared with the model control group, p<0.01). The MDA content of the combination of OPO and taurine (250+20.1 μg / mL) was the lowest, and was lower than the average value of the effects of the two alone at the same concentration (1.80<1.94). OPO and taurine had a synergistic effect at this ratio, and could achieve better results than the same concentration of monomers alone.

[0151] Table 3-1. Results of the evaluation experiment on the influence of samples on the content of malondialdehyde (MDA) (n=3)

[0152]

[0153] 3.2 Effects on CAT activity

[0154] Catalase (CAT) is an antioxidant enzyme widely found in organisms. It can catalyze the decomposition of hydrogen peroxide into water and oxygen, thereby protecting cells from oxidative damage caused by hydrogen peroxide. The test results show (as shown in Table 3-2 and Figure 8 As shown in the figure, the CAT activity in the model control group was significantly lower than that in the normal control group, indicating that the model was successfully established. Compared with the model control group, monomeric OPO (250μg / mL), monomeric taurine (20.1μg / mL), and the combination of OPO and taurine (250+20.1μg / mL) all increased the CAT activity in zebrafish eye cells, indicating that the three can play a certain protective role against blue light damage. Among them, monomeric OPO (250μg / mL) and the combination of OPO and taurine (250+20.1μg / mL) had a more significant effect on increasing CAT activity (compared with the model control group, p<0.05 or p<0.01). At the same time, the data showed that the CAT activity in zebrafish eye cells under the action of the combination (OPO and taurine, concentrations of 250 μg / mL and 20.1 μg / mL) was higher than the average value of the monomer effect under the same concentration conditions (Table 3-2, 0.574>0.456). OPO and taurine have a synergistic effect at this ratio, that is, the combination can produce an effect greater than the same concentration when the monomers are used separately.

[0155] Table 3-2. Results of the experiment evaluating the effect of samples on catalase (CAT) activity (n=3)

[0156]

[0157] Example 4: Evaluation of the effect on the expression of inflammation-related genes

[0158] 1. Test materials

[0159] 1.1. Sample preparation information

[0160] The solvent for OPO was DMSO, and the solvent for taurine was standard dilution water.

[0161] 1.2. Experimental animals

[0162] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), and were bred and provided by the Huante Biological Fish Farming Center. The experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0163] When conducting the experiment, the method of administering different samples to zebrafish was the same as described in Example 1.

[0164] 1.3. Instruments, consumables and reagents

[0165] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China); ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); fully automatic sample rapid grinding instrument (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); blue light instrument (50w450nm, China); high-speed refrigerated centrifuge (Heraeus Fresco17, ThermoFisher, Germany); fully automatic nucleic acid extraction instrument (Auto-Pure32A, Hangzhou Aosheng Instrument Co., Ltd., China); ordinary PCR amplifier (T100, BIO-RAD, Singapore); fluorescence quantitative PCR instrument (CFX Connect, BIO-RAD, Singapore); UV-visible spectrophotometer (Nanodrop2000, Thermo, USA); microplate mini centrifuge (BE-6100, Haimen Qilin Bell Instrument Manufacturing Co., Ltd., China); optical adhesive sealing film B (MSB1001, Bio-rad, USA); low-skirt 96-well plate (transparent) (HSP9601, Bio-rad, USA).

[0166] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); pronase E (Shanghai Yuanye Biotechnology Co., Ltd., China); anhydrous ethanol (Sinopharm Chemical Reagent Co., Ltd., China); ChamQ Universal SYBR qPCR Master Mix (Vazyme, China); FastKing cDNA First-Strand Synthesis Kit (Genomics-free) (Tiangen Biochemical Technology (Beijing) Co., Ltd., China); Pre-loaded magnetic bead method universal RNA extraction kit C (Foshan Aowei Biotechnology Co., Ltd. (ONREW), China).

[0167] 2. Detection Method

[0168] Randomly select 1dpf wild-type AB strain zebrafish and irradiate with blue light after membrane rupture to establish a zebrafish blue light eye damage model. At 3dpf, select model zebrafish with good development status and randomly distribute them in 6-well plates, and treat 30 zebrafish in each well (experimental group). Dissolve the samples with a certain concentration (concentrations are shown in Table 4-3), and set up a normal control group and a model control group at the same time. The volume of each well is 3mL, and three biological replicates are set in parallel. After 1 day of treatment at 28℃, the total RNA of each group of zebrafish was extracted using a fully automatic nucleic acid extractor, and the total RNA concentration and purity were determined using a UV-visible spectrophotometer. Take 2.00μg of total RNA from zebrafish samples, and synthesize 20.0μL of cDNA according to the instructions of the cDNA first-strand synthesis kit. The expression of β-actin, il-1β and tnf-α genes was detected by q-PCR. β-actin was used as an internal reference for gene expression to calculate the relative RNA expression of il-1β and tnf-α genes. The statistical results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0169] 3. Test results

[0170] 3.1. RNA extraction results and primer sequence information

[0171] At the end of the experiment, zebrafish total RNA was extracted, and the RNA concentration and A260 / A280 ratio (Table 4-1) were measured by UV-visible spectrophotometer. The A260 / A280 ratios were between 1.8 and 2.2, indicating that the extracted zebrafish total RNA was of good quality and could be used for subsequent q-PCR experiments. The primer sequences are shown in Table 4-2.

[0172] Table 4-1. Total RNA concentration and A260 / A280 ratio (n=3)

[0173]

[0174] Table 4-2. Primer sequence information

[0175]

[0176] 3.2. Effects on the expression of inflammation-related genes

[0177] Interleukin 1-β (IL-1β) is a key proinflammatory cytokine and one of the most important mediators of inflammation and host response to infection. It is involved in a variety of autoimmune inflammatory responses and a variety of cellular activities, including cell proliferation, differentiation, and apoptosis. Excessive production of IL-1β is associated with pathophysiological changes that occur in different disease states, and IL-1β production is often increased in inflammatory infected cells. Tumor necrosis factor α (TNF-α) is a potent inducer of inflammatory responses and a key regulator of innate immunity. It is a proinflammatory cytokine produced mainly by macrophages and monocytes, mediates a variety of inflammatory diseases, has a direct pathogenic effect, and induces the production of other inflammatory factors to exert tissue destructive effects.

[0178] The test results show (Table 4-3, Fig. 9 , 10 ), after blue light irradiation, the relative expression of proinflammatory factors il-1β and tnf-α genes in the model control group were significantly increased, indicating that blue light irradiation caused obvious inflammatory response in zebrafish eye cells. After the action of monomeric OPO, monomeric taurine, and the combination of OPO and taurine, the relative expression of il-1β gene was downregulated, among which the effect of the combination was more obvious (compared with the model control group, p<0.05), that is, the combination can significantly improve the inflammatory response of zebrafish eyes caused by blue light irradiation, and this effect is also significantly better than the average value of the effect when the monomer acts alone at the same concentration (0.668<0.873, p=0.05). OPO and taurine have a synergistic effect at this ratio, that is, when the two are used in combination, they can achieve an effect that cannot be achieved when the monomer acts alone at the same concentration. For the relative expression of tnf-α gene, monomeric OPO, monomeric taurine, and the combination of OPO and taurine all have a tendency to increase it, but this effect is not obvious.

[0179] Table 4-3. Results of the experimental evaluation of the effects of samples on the expression of inflammation-related genes (n=3)

[0180]

[0181] Example 5: Evaluation of the effect on ocular tissue pathology

[0182] 1. Test materials

[0183] 1.1. Sample preparation information

[0184] The solvents for lutein and OPO were DMSO, and the solvent for taurine was standard dilution water.

[0185] 1.2. Experimental animals

[0186] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), and were bred and provided by the Huante Biological Fish Farming Center. The experimental animal use license number is: SYXK (Zhejiang) 2022-0004. The breeding and management complies with the requirements of the international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0187] When conducting the experiment, the method of administering different samples to zebrafish was the same as described in Example 1.

[0188] 1.3. Instruments, consumables and reagents

[0189] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China); ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); blue light instrument (50w450nm, China); electronic constant temperature stainless steel water bath (HHS-2S, Shanghai Kanglu Instrument Equipment Co., Ltd., China); slicer (KD2258, Jinhua Kedi Instrument Equipment Co., Ltd., China); intelligent electric heating plate (400X280, Tianjin Leyuenag Laboratory Instrument Sales Co., Ltd., China); biological microscope (CX31, OLYMPUS, Japan).

[0190] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); pronase E (Shanghai Yuanye Biotechnology Co., Ltd., China); 4% tissue cell fixative (Beijing Solebow Technology Co., Ltd., China); anhydrous ethanol (Sinopharm Chemical Reagent Co., Ltd., China); xylene (Sinopharm Chemical Reagent Co., Ltd., China); Mayer hematoxylin staining solution (Shanghai Yihe Biotechnology Co., Ltd., China); eosin staining solution (Shanghai Yihe Biotechnology Co., Ltd., China); hydrochloric acid (Xilong Chemical Co., Ltd., China); neutral gum (Beijing Solebow Technology Co., Ltd., China); high-efficiency section paraffin (melting point 54-56°C, Shanghai Huayong Paraffin Co., Ltd., China); high-efficiency section paraffin (melting point 62-64°C, Shanghai Huayong Paraffin Co., Ltd., China).

[0191] 2. Detection Method

[0192] Wild-type AB zebrafish of 1dpf were randomly selected and irradiated with blue light to establish a zebrafish blue light eye injury model. Model zebrafish with good development at 3dpf were selected and randomly distributed in 6-well plates, with 30 zebrafish in each well (experimental group). Samples (concentrations see Fig.12 ), and set up a normal control group and a model control group at the same time, with a volume of 3 mL per well. After 1 day of treatment at 28°C, the zebrafish were fixed with 4% tissue cell fixative, and after a series of dehydration-embedding-sectioning-staining steps, the zebrafish were subjected to histopathological H&E staining analysis. The effects of the samples on the ocular histopathology of zebrafish were evaluated by ocular histopathology analysis.

[0193] 3. Test results

[0194] Fig.11 The zebrafish eye tissue analysis area slices and the names of each tissue are displayed. The test results show that ( Fig.12 ), the size and shape of the zebrafish eye tissue in the normal control group were normal, the retinal structure was clear and complete, and the cells at all levels were arranged neatly and orderly. Compared with the normal control group, the size of the zebrafish eyes in the model control group after blue light irradiation was reduced (such as Fig.12 The retinal structure is disordered, not hierarchical, and arranged in disorder (as shown in ① in the figure). Fig.12 The outermost protective layer, the pigment epithelium, becomes thinner and the pigment becomes diffuse, changing from black to lighter dark brown (as shown in Figure 2). Fig.12 The photoreceptor cone and rod cell layers are arranged irregularly (as shown in ③). Fig.12 The cells of the outer nuclear layer and inner nuclear layer, which play a structural supporting role, are arranged irregularly (as shown in Fig.12All the above signs indicate that the zebrafish blue light eye damage model was successfully established.

[0195] Compared with the model control group, the size of zebrafish eyes was significantly restored at a concentration of 1000 μg / mL of lutein, the retina was arranged in a hierarchical structure and in an orderly manner, the thickness of the pigment epithelium was significantly thickened without pigment diffusion, the cone and rod cell layers were arranged regularly, and the arrangement of cells in the outer nuclear layer and inner nuclear layer was restored. The size of zebrafish eyes was also significantly restored at the concentration of OPO and taurine combination (250+20.1 μg / mL), the retina was arranged in a hierarchical structure and in an orderly manner, the thickness of the pigment epithelium was significantly thickened without pigment diffusion, the cone and rod cell layers were arranged regularly, the cells in the outer nuclear layer and inner nuclear layer were arranged regularly, and the eye tissue morphology was similar to that of zebrafish in the normal control group. It can be judged that after the action of lutein (1000 μg / mL) and OPO and taurine combination (250+20.1 μg / mL), the tissue structure of the zebrafish eyes irradiated by blue light was restored, and they have a mitigating and improving effect on the damage to the zebrafish eye tissue caused by blue light irradiation.

[0196] In summary, blue light has high energy, and a certain degree of blue light exposure will cause oxidative damage to the zebrafish eye, which is specifically manifested as abnormal accumulation of reactive oxygen free radicals (ROS) in the eye, and a decrease in the activity of eye-related antioxidant enzymes (such as catalase, CAT), which in turn leads to an increase in lipid oxidation in the eye, and lipid oxidation-related products such as malondialdehyde (MDA) will increase accordingly. Oxidative damage leads to an increase in the relative expression of eye proinflammatory factors such as IL-1β and TNF-α genes il-1β and tnf-α, inducing an inflammatory response, and ultimately leading to apoptosis of zebrafish eye cells and even abnormal changes in tissue structure. This experiment found that a certain concentration of monomeric OPO, monomeric taurine, and a certain proportion of the combination of OPO and taurine can alleviate the damage caused by blue light irradiation, specifically reducing the degree of apoptosis of eye cells, removing active oxygen free radicals in the eye, reducing the content of lipid oxidation products (MDA), increasing the activity of antioxidant enzymes (CAT), reducing the relative expression of the proinflammatory factor IL-1β gene il-1β, inhibiting the occurrence of inflammation, and improving the structure and state of damaged eye tissue. By comparison, it was found that a certain proportion of OPO+taurine composition (500:0.403~40.2) can reduce the apoptosis of eye cells caused by blue light irradiation to a certain extent. And the effect of the combination is better than the average effect of the same concentration of monomers used alone, which means that these compositions have a synergistic effect, and when used in combination, they can achieve an effect that cannot be achieved when the monomers are used alone. In particular, when the combined concentration of the two is 250+20.1 μg / mL (the ratio of the two is 500:40.2), although its total concentration (270.1 μg / mL) is lower than the total concentrations when the concentrations of the two are 500+0.403, 500+1.34, 500+6.71 μg / mL (the ratio of the two is 500:(0.403-6.71)), the composition has a very obvious anti-apoptosis effect on ocular cells, and its effect is better than the effect of OPO concentration of 250 μg / mL and taurine concentration of 20.1 μg / mL when used alone (p<0.05), and is also significantly better than the average effect of the two when used alone (p<0.05). Therefore, it can be considered that the effect of the two at this ratio (500:40.2) is the best, and the synergistic effect is most significant. Taking this combination as an example, it was further confirmed that the combination of OPO and taurine can effectively remove the reactive oxygen free radicals generated by blue light irradiation in zebrafish eyes, reduce the content of lipid oxidation products (MDA) in the eyes, increase the activity of antioxidant enzymes (CAT), reduce the relative expression of the pro-inflammatory factor IL-1β gene il-1β, inhibit the occurrence of inflammation, and improve the structure and state of damaged eye tissues. And the above effects of the combination are better than the average effect of the monomer components used alone at the same concentration, so it has a synergistic effect.

[0197] Industrial Applicability

[0198] The composition provided by the invention can be produced and used on an industrial scale.

Claims

1. A composition, characterized in that The composition comprises the following essential components: 1,3-dioleyl-2-palmitoylglycerol and taurine; Furthermore, in the composition, the mass ratio of the 1,3-dioleyl-2-palmitoylglycerol to the taurine is 500:(0.38-13.42).

2. Use of the composition according to claim 1 in preparing food that helps alleviate blue light damage.

3. The use according to claim 2, characterized in that: The method helps to alleviate blue light damage, including at least one of reducing eye cell apoptosis, reducing the accumulation of active oxygen free radicals in the eye, reducing the content of lipid oxidation products in the eye, increasing the activity of antioxidant enzymes in the eye, reducing the expression of the gene il-1β and improving the eye tissue structure.

4. The use according to claim 3, characterized in that The lipid oxidation products include malondialdehyde.

5. The use according to claim 3, characterized in that: The antioxidant enzymes include catalase.

6. The use according to claim 3, characterized in that: The improvement of eye tissue structure includes at least one of restoring the eye size, arranging the retina in an orderly hierarchical structure, thickening the pigment epithelium without pigment diffusion, arranging the cone and rod cell layers in a regular manner, and arranging the outer nuclear layer and inner nuclear layer cells in a regular manner.

7. The use according to any one of claims 2 to 6, characterized in that Under normal temperature conditions, the food is a solid dairy product or a liquid dairy product.

8. The use according to any one of claims 2 to 6, characterized in that: The food is infant food or children's food.

9. The use according to any one of claims 2 to 6, characterized in that: The food further contains any one or more of the following ingredients: plant product ingredients, animal dairy product ingredients, animal meat product ingredients, functional added ingredients and any acceptable auxiliary materials.

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