A composition to reduce blue light damage

Abstract

This invention belongs to the field of functional nutrient research technology, specifically relating to a composition for reducing blue light damage. The functional composition provided by this invention comprises the following essential components: taurine and coenzyme Q10; and, in the functional composition, the effective ratio of taurine to coenzyme Q10 is (0.1–8):1. Compared to taurine monomers or coenzyme Q10 monomers, the composition provided by this invention can more effectively reduce the damage to the eyes caused by blue light irradiation, especially by scavenging reactive oxygen species generated by blue light irradiation, reducing the content of ocular lipid oxidation products such as malondialdehyde (MDA), increasing the activity of antioxidant enzymes such as catalase, reducing the relative expression of the pro-inflammatory factor IL-1β gene il-1β, inhibiting the occurrence of inflammation, and improving the structure and condition of damaged ocular tissues.

Description

Technical Field

[0001] This invention belongs to the field of functional nutrient research technology, specifically relating to a composition that reduces blue light damage. Background Technology

[0002] Blue light, with wavelengths between 400 nm and 500 nm, is visible light and an important component of natural light. Blue light has certain physiological functions, helping to maintain normal vision and human health, participating in visual perception and color vision processes, and involved in pupillary light reflex and pupillary constriction. It also plays a role in regulating the biological clock. However, blue light has high energy and strong penetrating power, reaching directly to the retina and causing blue light damage. The main mechanism of blue light damage is oxidative damage. Blue light stimulates the retina to initiate a photo-oxidation mechanism, inducing mitochondria to produce a large number of free radicals, leading to lipid peroxidation, messenger RNA damage, and protein destruction. This results in apoptosis of eye cells such as photoreceptor cells and pigment epithelial cells, causing degenerative changes in the retina. In severe cases, it can lead to vision impairment, macular degeneration, and symptoms such as red eyes, dry eyes, eye strain, blurred vision, and eye fatigue. It can also cause eye diseases such as cataracts. Besides naturally occurring blue light, artificial light sources such as light-emitting diodes (LEDs) are also a significant source of blue light. LEDs primarily emit white light by exciting yellow phosphors with blue light-emitting chips. Therefore, at high color temperatures, the light source spectrum exhibits a strong blue light peak. With the rapid development of LED-related technologies, they are widely used in daily life, especially in various screen electronic products. However, due to the significant changes in work patterns, people rely heavily on mobile phones, tablets, and computers for daily work, communication, and entertainment, spending hours or even tens of hours a day using these electronic devices. This prolonged exposure to blue light significantly increases the risk of various eye diseases.

[0003] Currently, blue light protection primarily includes physical and chemical methods. Physical protection mainly involves using blue light blocking glasses, screens, and films to physically block blue light. By blocking some blue light from reaching the eyes, physical methods can alleviate eye strain from using electronic devices and protect the retina from blue light damage. However, they also interfere with the positive effects of blue light on normal visual function and physiological regulation. Chemical protection methods mainly include taking antioxidants, free radical scavengers, and even anti-inflammatory drugs. Because retinal cells and lens epithelial cells produce free radicals such as singlet oxygen or superoxide anions when exposed to blue light, this causes oxidative stress on eye cells and tissues. Antioxidants or free radical scavengers can inhibit the accumulation of oxidative stress. For example, lutein can protect cells from photo-oxidation and photodamage, acting as an active high-energy blue light filter to provide antioxidant protection for organisms; vitamin E can, to some extent, scavenge reactive oxygen species (ROS) generated by blue light irradiation; vitamin C can also, to some extent, prevent cellular oxidative damage, thereby preventing blue light irradiation from damaging mitochondrial DNA. While blue light irradiation leads to the production and accumulation of ROS, it can also promote the release of inflammatory factors; therefore, anti-inflammatory drugs can, to some extent, alleviate blue light damage. Chemical protection methods do not interfere with the normal physiological functions of blue light and can specifically alleviate blue light damage.

[0004] Taurine is a stable endogenous sulfur-containing amino acid found in living organisms. It is widely distributed in various tissues and organs of humans and animals, with higher concentrations in skeletal muscle and the heart. Furthermore, taurine is present in mammalian milk and is an essential nutrient for infant growth and development. Studies have shown that taurine, as a conditionally essential amino acid, possesses various biological functions, including enhancing immunity, relieving fatigue, anti-oxidation, promoting fat metabolism, protecting the nervous system, and improving memory. Reference 1 discloses a health food product for relieving eye fatigue and its preparation method. This health food product composition contains taurine, zinc gluconate, lutein, bilberry extract, vitamin E, and vitamin C. However, it does not provide empirical evidence regarding the effectiveness of this composition in relieving eye fatigue. Reference 2 discloses a composition for an anti-blue light eye spray, comprising 0.05-0.1% dried orange peel 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, and 0.05-0.2% taurine. However, it does not provide specific effects of the above composition on resisting the harmful effects of blue light on the eyes.

[0005] Coenzyme Q10 is a lipid-soluble quinone compound composed of 10 isopentenyl units. It is mainly found on the inner mitochondrial membrane and participates in respiratory chain electron transport, antioxidation, metabolic regulation, and cell differentiation regulation. It is a natural antioxidant and free radical scavenger. Reduced coenzyme Q10 has strong antioxidant function and can work synergistically with vitamin E, ascorbic acid, and other substances to eliminate excess free radicals in the body and protect cells from oxidative stress damage. Reference 3 discloses a compound preparation with anti-radiation and vision-protecting functions. The formula, by weight, is 65-98% tea seed oil, 0.15-15.0% coenzyme Q10, 0.05-10.0% lutein, 0.10-10.0% alpha-lipoic acid, and 0.01-0.10% vitamin A. It claims that the combination of these five components forms a highly effective anti-radiation and vision-protecting product. However, it does not provide empirical evidence for the actual efficacy of this combination. Reference 4 discloses a composition for enhancing human anti-fatigue, comprising the following components: 2-15 kg of coenzyme Q10, 2-12 kg of conjugated linoleic acid, 5-25 kg of D-ribose, 0.15-2 kg of octacosanol, 2-12 kg of L-carnitine, 2-12 kg of calcium pyruvate, 2-15 kg of ginseng, 3-20 kg of codonopsis pilosula, 3-20 kg of pseudostellaria heterophylla, 5-25 kg of astragalus membranaceus, 2-15 kg of daidzein, 0-20 kg of coptis chinensis, 0-20 kg of rhubarb, 2-12 kg of cinnamon, 2-15 kg of chicken gizzard lining, 2-15 kg of hawthorn, 5-20 kg of raw materials for relieving eye fatigue, 5-20 kg of antioxidants, and 5-25 kg of amino acids. The raw materials for relieving eye fatigue are black soybean anthocyanins, taurine, D-linolenic acid, lutein, kelp extract, and mulberry leaf extract, mixed in a 1:1…1 ratio. Although the above composition contains ingredients that claim to relieve eye strain, the composition as a whole is a composition that enhances the body's resistance to fatigue, rather than a composition that relieves eye strain, and it does not provide empirical evidence that the above composition relieves eye strain.

[0006] References:

[0007] Reference 1: CN108771243A;

[0008] Reference 2: CN109549947A;

[0009] Reference 3: CN101670019B;

[0010] Reference 4: CN115554387A. Summary of the Invention

[0011] The problem the invention aims to solve

[0012] Although existing technologies have researched nutrients that are beneficial to eye health, such as relieving eye fatigue, protecting against blue light, resisting radiation, or protecting vision, such research cannot be considered perfect, and there is still room for development of chemical protection methods to reduce blue light damage.

[0013] In this regard, the present invention aims to provide a functional composition for reducing blue light damage using taurine and coenzyme Q10. By constructing a zebrafish model of blue light damage, the present invention tested the effects of taurine and coenzyme Q10 alone, as well as combinations thereof, on reducing blue light damage. Unexpectedly, it was found that when taurine and coenzyme Q10 are combined in a certain proportion, the effect that cannot be achieved when they are used alone can be obtained. That is, the present invention found that taurine and coenzyme Q10 have a synergistic effect in reducing blue light damage.

[0014] Solution for solving the problem

[0015] To address the aforementioned technical problems, the present invention provides the following technical solution:

[0016] A first aspect of the present invention provides a functional composition comprising the following essential components:

[0017] Taurine and Coenzyme Q10;

[0018] Furthermore, in the functional composition, the effective ratio of taurine to coenzyme Q10 is (0.1~8):1.

[0019] In some embodiments, the effective ratio of taurine to coenzyme Q10 in the functional composition is (1.5~7.5):1.

[0020] In some implementations, the benefits of mitigating blue light damage include at least one of reducing the accumulation of reactive oxygen species in the eye, reducing the content of lipid peroxidation products in the eye, increasing the activity of antioxidant enzymes in the eye, reducing the expression of the il-1β gene, reducing ocular cell apoptosis, and improving ocular tissue structure.

[0021] In some embodiments, reducing ocular lipid peroxidation product levels includes reducing ocular malondialdehyde levels.

[0022] In some embodiments, the enhancement of ocular antioxidant enzyme activity includes enhancing ocular catalase activity.

[0023] In some implementations, the improvement of ocular tissue structure includes at least one of restoring eye size, promoting a hierarchical and orderly arrangement of the retina, promoting thickening of the pigment epithelium without pigment diffusion, promoting regular arrangement of cone and rod cell layers, and promoting regular arrangement of outer and inner nuclear cell layers.

[0024] The effects of the invention

[0025] By implementing the above technical solution, the present invention achieves the following technical effects:

[0026] This invention provides a functional composition comprising taurine and coenzyme Q10 in a specific ratio. Experimental data show that, compared to taurine monomers or coenzyme Q10 monomers, the functional composition provided by this invention can more effectively reduce eye damage caused by blue light irradiation. In particular, it can scavenge reactive oxygen free radicals generated by blue light irradiation, reduce the content of lipid oxidation products in the eye such as malondialdehyde, increase the activity of antioxidant enzymes such as catalase, reduce the relative expression level of the pro-inflammatory factor IL-1β gene il-1β, inhibit the occurrence of inflammation, and improve the structure and condition of damaged eye tissues. Attached Figure Description

[0027] Figure 1 Example 1: Fluorescence intensity of apoptotic cells in the eyes of zebrafish after sample treatment; where, compared with the model control group, **p<0.01, ***p<0.001.

[0028] Figure 2 Example 1: Typical fluorescence intensity of apoptotic cells in the eyes of zebrafish after sample treatment; where the fluorescent particles indicated by the arrows are apoptotic cells.

[0029] Figure 3 Example 1: Fluorescence intensity of apoptotic cells in the eyes of zebrafish after sample treatment; Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001; compared with coenzyme Q10 40.0 μg / mL, .

[0030] Figure 4 Example 1: Typical fluorescence intensity of apoptotic cells in the eyes of zebrafish after sample treatment; where the fluorescent particles indicated by the arrows are apoptotic cells.

[0031] Figure 5 Example 2: Average fluorescence intensity of zebrafish heads after sample treatment; compared with the model control group, *p<0.05, ***p<0.001; compared with taurine concentration of 250 μg / mL, ^ p < 0.05; compared with a coenzyme Q10 concentration of 40.0 μg / mL, .

[0032] Figure 6 Example 2: Typical graph of average fluorescence intensity of zebrafish head after sample treatment; the area within the dashed box is the analysis area of ​​zebrafish head.

[0033] Figure 7Example 3: MDA content in zebrafish after sample treatment; where, compared with the model control group, *p<0.05, **p<0.01, ***p<0.001.

[0034] Figure 8 Example 3: CAT activity in zebrafish after sample treatment; where, compared with the model control group, *p<0.05, **p<0.01, ***p<0.001.

[0035] Figure 9 Example 4: Relative expression level of zebrafish il-1β gene after sample treatment; where, compared with the model control group, *p<0.05.

[0036] Figure 10 Example 5: Zebrafish eye tissue analysis area and cross-sectional retinal slice; where the black single-sided square brackets represent the zebrafish retina.

[0037] Figure 11 Example 5: Typical diagram of zebrafish eye tissue structure after sample treatment; where ① is a white double-arrowed cross, representing the sum of the anterior-posterior axis (A / P) and dorsoventral axis (D / V) dimensions of the eye; ② is a black half-square bracket, representing the retinal structure; ③ is a black arrow, representing the pigment epithelium; ④ is a white dashed elliptical area, representing the cone and rod cell layers; ⑤ is a black dashed fan-shaped area, representing the outer nuclear layer and inner nuclear layer. Detailed Implementation

[0038] The following describes embodiments of the present invention, but the present invention is not limited thereto. The present invention is not limited to the various configurations described below, and various modifications can be made within the scope of protection claimed by the present invention. 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.

[0039] I. Terminology Definition

[0040] In this invention, the range of values ​​represented by “value A ~ value B”, “value A - value B”, “value A or above” or “value A or below” refers to the range that includes the endpoint values ​​A and B.

[0041] In this invention, the terms "comprising," "including," or "containing" can mean included or open-ended, and do not exclude additional, uncited components or method steps. At the same time, "comprising," "including," or "containing" can also mean closed-ended, excluding additional, uncited components or method steps.

[0042] In this invention, the term "about" is used to define that the numerical ranges and parameters of this invention are approximate values, while specific related values ​​have been presented as precisely as possible. Unless otherwise explicitly stated, it should be understood that all ranges, quantities, values, and percentages used in this invention are modified by "about". Here, "about" generally means that the actual value is within ±3%, ±2%, ±1%, or ±0.5% of a specific value or range.

[0043] In this invention, "young adults" refers to a group of people aged 18 years or older and less than 45 years old.

[0044] In this invention, "middle-aged and elderly" refers to the human population aged 45 years and older.

[0045] In this invention, the term "normal temperature" or "room temperature" refers to an indoor ambient temperature of "23±2℃".

[0046] In this invention, the term "animal milk" is used to refer to the fluid obtained from the mammary glands of a mammal in the process of lactation. The term "animal milk" should be interpreted broadly and encompasses both raw milk (i.e., the fluid obtained directly from the mammary glands) and standardized dairy products (such as skim milk or whole milk).

[0047] Unless otherwise defined, other technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0048] II. Functional Compositions

[0049] This invention provides a functional composition comprising the essential components of taurine and coenzyme Q10; and wherein, in the functional composition, the effective ratio of taurine to coenzyme Q10 is (0.1~8):1; exemplary ratios may be approximately 0.15:1, 0.17:1, 0.18:1, 0.19:1, 0.2:1, 0.3:1, 0.4:1, 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1.0:1, 1.3:1, 1.6:1, 1.9:1, 2.0:1, 2.3:1, 2.6:1, 2.9:1, 3.0:1, 3.3:1, 3.6:1, 3.9:1, 4.0:1, 4.3:1. 1, 4.6: 1, 4.9: 1, 5.0: 1, 5.3: 1, 5.6: 1, 5.9: 1, 6.0: 1, 6.3: 1, 6.6: 1, 6.9: 1, 7.0: 1, 7.1: 1, 7.14: 1, 7.2: 1, 7.5: 1, 7.7: 1 or 7.9: 1, etc.; preferably an effective ratio of (0.17~7.5):1, more preferably an effective ratio of (1.5~7.5):1, even more preferably an effective ratio of (2.00~7.14):1, and even more preferably an effective ratio of (6.0~6.5):1.

[0050] Compared to taurine monomers or coenzyme Q10 monomers, compositions containing taurine and coenzyme Q10 have shown greater effectiveness in reducing blue light damage to the eyes, especially when the ratio of taurine to coenzyme Q10 is in the range of (2.00~7.14):1.

[0051] This invention does not impose particular limitations on the preparation method or source of taurine and coenzyme Q10. For example, taurine can be prepared by biological extraction, chemical synthesis (such as esterification, chlorination, etc.), and coenzyme Q10 can be prepared by microbial fermentation, biological extraction, cell culture, chemical synthesis, etc. In some embodiments, this invention preferably uses oxidized coenzyme Q10.

[0052] In some embodiments, the functional composition comprises taurine and coenzyme Q10, and the effective ratio of taurine to coenzyme Q10 in the functional composition is (0.1~8):1.

[0053] The present invention does not particularly limit the form of the functional composition. For example, the functional composition may be a liquid or a solid at room temperature.

[0054] The functional composition described in this invention is a synthetic or compounded composition, and not a naturally occurring composition.

[0055] In some embodiments, in the functional composition of the present invention, the effective amount of taurine is greater than or equal to 6.71 µg / mL, preferably greater than or equal to 20.1 µg / mL, more preferably greater than or equal to 80 µg / mL, and even more preferably 80~250 µg / mL; the effective amount of coenzyme Q10 is greater than or equal to 30 µg / mL, preferably greater than or equal to 35 µg / mL, and more preferably 35~40 µg / mL.

[0056] III. Uses to reduce blue light damage

[0057] This invention is the first to propose a specific ratio of taurine and coenzyme Q10 to effectively reduce the damage to the eyes caused by blue light exposure, and taurine and coenzyme Q10 have a synergistic effect. Furthermore, the reduction of blue light damage described in this invention is not intended to treat or prevent disease, and the blue light damage described does not reach the level of a disease.

[0058] In some implementations, the benefits of mitigating blue light damage include at least one of reducing the accumulation of reactive oxygen species in the eye, reducing the content of lipid peroxidation products in the eye, increasing the activity of antioxidant enzymes in the eye, reducing the expression of the il-1β gene, reducing ocular cell apoptosis, and improving ocular tissue structure.

[0059] In some specific implementations, reducing the content of ocular lipid oxidation products includes reducing the content of ocular malondialdehyde.

[0060] In some specific implementations, the enhancement of ocular antioxidant enzyme activity includes enhancing ocular catalase activity.

[0061] In some specific implementations, the improvement of ocular tissue structure includes at least one of the following: restoring eye size, promoting orderly hierarchical arrangement of the retina, thickening of the pigment epithelium without pigment diffusion, promoting regular arrangement of cone and rod cell layers, and promoting regular arrangement of outer and inner nuclear cell layers.

[0062] Example

[0063] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified, specific conditions in the examples are performed under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, all materials and instruments used are commercially available conventional products.

[0064] Example 1: Evaluation of anti-ocular apoptosis efficacy

[0065] 1. Experimental materials

[0066] 1.1. Sample Preparation Information

[0067] Taurine was purchased from DSM (China) Co., Ltd., and the solvent was standard dilution water.

[0068] Coenzyme Q10 (oxidized form) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., and the solvent was DMSO.

[0069] Positive control: Lutein, purchased from DSM (China) Co., Ltd., in DMSO solvent.

[0070] 1.2. Laboratory Animals

[0071] Zebrafish were raised in aquarium water at 28℃ (water quality: 200 mg of readily soluble sea salt added per 1 L of reverse osmosis water; conductivity 450~550 μS / cm; pH 6.5~8.5; hardness 50~100 mg / L CaCO3), bred and provided by Hangzhou Huante Biological Fish Farming Center. The laboratory animal use license number is: SYXK (Zhejiang) 2022-0004. The husbandry and management met the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0072] During the experiment, zebrafish were given taurine monomer as follows: taurine was dissolved in standard dilution water to the target concentration, and the zebrafish were then placed in the sample solution of the target concentration. Zebrafish were given coenzyme Q10 monomer as follows: coenzyme Q10 was first dissolved in DMSO as a stock solution, and then the stock solution was diluted with standard dilution water to the target concentration using the miscibility of DMSO and water. The zebrafish were then placed in the sample solution of the target concentration. The method for giving zebrafish lutein monomer was the same as that for giving zebrafish coenzyme Q10 monomer. The method for giving zebrafish a combination of coenzyme Q10 and taurine was as follows: coenzyme Q10 was first dissolved in DMSO as a stock solution, and then the coenzyme Q10 stock solution was diluted with standard dilution water using standard dilution water while simultaneously dissolving taurine to the target concentration using the miscibility of DMSO and water. The zebrafish were then placed in the sample solution of the target concentration.

[0073] 1.3. Instruments, Consumables and Reagents

[0074] 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 Belamber Biotechnology Co., Ltd., China); Motorized focusing continuous zoom fluorescence microscope (AZ100, Nikon, Japan); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); Fully automatic rapid sample grinder (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); Blue light meter (50W 450nm, China).

[0075] Dimethyl sulfoxide (DMSO, Sigma, Switzerland; I2229063, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Streptase E (Shanghai Yuanye Biotechnology Co., Ltd., China); Acridine orange (AO, Shanghai Maclean Biotechnology Co., Ltd., China); Methylcellulose (Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0076] 2. Test Methods

[0077] 2.1. Determination of Maximum Detectable Concentration (MTC)

[0078] Wild-type AB strain zebrafish were randomly selected one day after fertilization (1 dpf) and exposed to blue light to establish a zebrafish blue light eye damage model. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water (concentrations shown in Table 1-1), and a normal control group and a model control group were also included. The volume of each well was 3 mL. After treatment at 28℃ for 1 day, the MTC of the samples in the model zebrafish was measured.

[0079] 2.2. Evaluation of anti-ocular apoptosis efficacy

[0080] 2.2.1 Evaluation of the anti-apoptotic efficacy of the monomeric components

[0081] Wild-type AB strain zebrafish at 1 dpf were randomly selected and irradiated with blue light to establish a zebrafish blue light-induced eye damage model. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Different concentrations of samples were administered in water (see Tables 1-2). A positive control of lutein concentration of 1000 μg / mL was set up, along with a normal control and a model control group. Each well had a volume of 3 mL. After treatment at 28℃ for 1 day, zebrafish in each experimental group underwent AO staining in the dark for 30 min. After washing three times with standard dilution water, 10 zebrafish from each experimental group were randomly selected and photographed under a fluorescence microscope. ImageJ software was used to analyze and collect data. The fluorescence intensity of apoptotic cells in the zebrafish eyes was analyzed and statistically analyzed. The anti-apoptotic efficacy of the samples was evaluated based on the statistical analysis results of the above indicators. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. p < 0.05 indicated statistical significance.

[0082] 2.2.2 Evaluation of the anti-ocular apoptosis efficacy of the composition

[0083] Based on the evaluation results of the anti-apoptotic efficacy of the above-mentioned monomeric components, a combination experiment was conducted using optimal concentrations of taurine and coenzyme Q10 to evaluate the relevant effects of the combination. The experimental conditions were the same as the evaluation methods described above. A positive control lutein concentration of 1000 μg / mL was set up, along with a normal control group and a model control group. After the experiment, the zebrafish treatment, data collection, and analysis methods were the same as described above.

[0084] 3. Experimental Results

[0085] 3.1. MTC

[0086] The experimental results showed that the MTC of positive controls lutein and taurine in model zebrafish was 2000 μg / mL, while the MTC of coenzyme Q10 in model zebrafish was 250 μg / mL, as detailed in Table 1-1.

[0087] Table 1-1. Results of the concentration exploration experiment for the anti-apoptotic efficacy of samples (n = 30)

[0088]

[0089] 3.2. Evaluation of anti-ocular apoptosis efficacy

[0090] 3.2.1 Evaluation of the anti-apoptotic efficacy of the monomeric components

[0091] The experimental results showed that after 1 dpf wild-type AB strain zebrafish were irradiated with blue light, the fluorescence intensity of apoptotic cells in their eyes increased significantly (Table 1-2). Figure 1The blue light irradiation significantly increased apoptosis in zebrafish eye cells, indicating successful model establishment. Meanwhile, the positive controls lutein, taurine, and coenzyme Q10 all showed significant anti-apoptotic effects at certain concentrations (p < 0.01 or 0.001 compared to the fluorescence intensity of apoptotic cells in the model control group), specifically at lutein 1000 μg / mL, taurine 1000 and 2000 μg / mL, and coenzyme Q10 62.5, 125, and 250 μg / mL (see Tables 1-2). The fluorescence intensity of apoptotic cells in zebrafish eye cells was also observed. Figure 2 It can also be observed that, compared with the model control group, the fluorescence intensity / spot size of apoptotic cells treated with the above-mentioned components tended to decrease / become smaller, and the higher the concentration of the monomeric component, the more obvious the above effect was.

[0092] Table 1-2. Experimental results evaluating the anti-apoptotic efficacy of monomeric components (n = 10)

[0093]

[0094] 3.2.2 Evaluation of the anti-ocular apoptosis efficacy of the composition

[0095] Based on the above evaluation results of the anti-apoptotic efficacy of the monomeric components, we selected different concentrations of taurine and coenzyme Q10 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 2018. Figure 3 Under the conditions of this experiment, we found that neither taurine nor coenzyme Q10 monomers at the selected concentrations significantly reduced the fluorescence intensity of zebrafish eye cells, and therefore did not have a significant effect in preventing blue light damage. However, when taurine and coenzyme Q10 were combined, although the concentrations of both were still below the effective concentrations of the monomers, some combinations showed significant anti-apoptotic effects in zebrafish eyes. Furthermore, the fluorescence intensity of apoptotic zebrafish eye cells under the combined conditions was lower than the average fluorescence intensity of cells when either taurine or coenzyme Q10 was used alone at the same concentration. The different ratios of taurine and coenzyme Q10 selected in this experiment (ratio of (0.17~7.14):1) all showed better effects than the average effect when either taurine or coenzyme Q10 was used alone at the same concentration, indicating a synergistic effect. When the concentrations of the taurine and coenzyme Q10 combination were 80.0 ± 40.0, 250 ± 35.0, and 250 ± 40 µg / mL, i.e., the ratio was (2.00–7.14):1, the fluorescence intensity of apoptotic cells in zebrafish eyes all decreased to very low levels (e.g., Figure 3As shown in the figure), compared with the model control group, p < 0.01 or 0.001, close to the level of the positive control lutein, and the fluorescence intensity was lower than the average effect of the two when used alone at the same concentration (p = 0.075, 0.065 and 0.045, the difference reached or approached the significant level), and the size and / or intensity of the fluorescent spots shown in the typical fluorescence intensity diagram showed a trend of decreasing and / or weakening (e.g. Figure 4 As shown in the figure, this indicates that the combination of the two at the above ratio has a good effect on preventing blue light-induced ocular cell apoptosis and has a significant synergistic effect. In particular, when the concentration of the combination is 250 ± 40.0 µg / mL (6.25:1), the composition has a very obvious effect on preventing ocular cell apoptosis, which is better than the effect of using coenzyme Q10 at a concentration of 40.0 µg / mL alone. Moreover, the effect of this composition is significantly better than the average effect of using either one alone (p < 0.05). It can be considered that the synergistic effect of the two at this concentration and ratio of 250 ± 40 µg / mL (6.25:1) is the most significant.

[0096] Table 1-3. Experimental results evaluating the anti-apoptotic efficacy of the composition (n = 10)

[0097]

[0098] Example 2: Evaluation of ROS removal effect

[0099] 1. Testing materials

[0100] 1.1. Sample Preparation Information

[0101] Taurine was diluted with standard water, coenzyme Q10 was diluted with DMSO, and the positive control lutein was also diluted with DMSO.

[0102] 1.2. Laboratory Animals

[0103] Zebrafish were raised in aquarium water at 28℃ (water quality: 200 mg of readily soluble sea salt added per 1 L of reverse osmosis water; conductivity 450~550 μS / cm; pH 6.5~8.5; hardness 50~100 mg / L CaCO3), bred and provided by Hangzhou Huante Biological Fish Farming Center. The laboratory animal use license number is: SYXK (Zhejiang) 2022-0004. The husbandry and management met the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0104] During the experiment, zebrafish were given different samples in the same manner as described in Example 1.

[0105] 1.3. Instruments, Consumables and Reagents

[0106] 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 Belamber Biotechnology Co., Ltd., China); Motorized focusing continuous zoom fluorescence microscope (AZ100, Nikon, Japan); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); Blue light meter (50W 450nm, China).

[0107] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Streptase E (Shanghai Yuanye Biotechnology Co., Ltd., China); Cell ROX Green Reagent (Invitrogen, USA); Methylcellulose (Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0108] 2. Detection Method

[0109] A zebrafish strain of the Albino strain with a melanin allele mutation was randomly selected at 1 dpf (day post-exposure) and exposed to blue light to establish a zebrafish blue light-induced eye damage model. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples and coenzyme Q10 (concentrations shown in Table 2-1) were administered in water, and a normal control group and a model control group were also included. Each well contained 3 mL of sample. After treatment at 28℃ for 1 day, zebrafish in each experimental group were stained with CellROX in the dark for 30 min. After washing three times with standard dilution water, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. Data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The average fluorescence intensity of the zebrafish head was analyzed and statistically analyzed to evaluate the effect of the sample on ROS levels. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. A p < 0.05 indicated that the difference was statistically significant.

[0110] 3. Test Results

[0111] Blue light irradiation leads to the accumulation of reactive oxygen species (ROS) in ocular cells, causing oxidative damage. Experimental results showed that after blue light irradiation, the ROS fluorescence intensity 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 ROS accumulation in the zebrafish eyes, and the model was successfully established. Compared with the model control group, the positive controls lutein (1000 μg / mL), taurine (250 μg / mL), coenzyme Q10 (40.0 μg / mL), and the taurine + coenzyme Q10 combination (250 + 40.0 μg / mL) all exhibited ROS scavenging effects, specifically by reducing the average fluorescence intensity in the zebrafish head (Table 2-1). Figure 5 The fluorescence levels in the zebrafish heads were significantly lower than those in the model control group (p < 0.05 or 0.001). Figure 6 Compared to the monomeric components taurine and coenzyme Q10, the composition (250 μg / mL taurine + 40.0 μg / mL coenzyme Q10, i.e., a ratio of 6.25:1) showed better scavenging of ROS generated by blue light irradiation (fluorescence value 2.14, p < 0.05 compared to 250 μg / mL taurine and 40.0 μg / mL coenzyme Q10). Furthermore, the composition's effect was significantly better than the average effect of the two monomers at the same concentration (p < 0.01). This demonstrates that taurine and coenzyme Q10 have a significant synergistic effect when used in this ratio, significantly enhancing the monomer's ability to scavenge ROS generated by blue light irradiation.

[0112] Table 2-1. Results of the ROS removal effect evaluation test on samples (n = 10)

[0113]

[0114] Example 3: Evaluation of the effect on the content / activity of oxidative damage-related enzymes

[0115] 1. Testing materials

[0116] 1.1 Sample Configuration Information

[0117] Taurine is dissolved in standard dilution water, and coenzyme Q10 is dissolved in DMSO.

[0118] 1.2 Experimental Animals

[0119] Zebrafish were raised in aquarium water at 28℃ (water quality: 200 mg of readily soluble sea salt added per 1 L of reverse osmosis water; conductivity 450~550 μS / cm; pH 6.5~8.5; hardness 50~100 mg / L CaCO3), bred and provided by Hangzhou Huante Biological Fish Farming Center. The laboratory animal use license number is: SYXK (Zhejiang) 2022-0004. The husbandry and management met the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0120] During the experiment, zebrafish were given different samples in the same manner as described in Example 1.

[0121] 1.3 Instruments, Consumables and Reagents

[0122] 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 Beilanbo Biotechnology Co., Ltd., China); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); Blue light analyzer (50W 450nm, China); 96-well plate (Nest Biotech, China); Fully automatic sample grinder (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); High-speed refrigerated centrifuge (Heraeus Fresco17, ThermoFisher, Germany); Multifunctional microplate reader (SPARK, TECAN, Austria); Electronic thermostatic stainless steel water bath (HHS-2S, Shanghai Kanglu Instrument Equipment Co., Ltd., China).

[0123] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Streptase E (Shanghai Yuanye Biotechnology Co., Ltd., China); BCA protein concentration assay kit (Boster Biological, 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).

[0124] 2. Detection Method

[0125] 2.1 Effect on malondialdehyde (MDA) content

[0126] Wild-type AB strain zebrafish at 1 dpf were randomly selected and irradiated with blue light to establish a zebrafish blue light eye damage model. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water (concentrations shown in Table 3-1), and a normal control group and a model control group were also set up. Each well contained 3 mL, and three biological replicates were performed. After treatment at 28℃ for 1 day, zebrafish samples were collected according to the MDA kit instructions, and MDA content was detected using a multi-mode microplate reader. The effect of the sample on MDA content was evaluated using statistical analysis. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software; p < 0.05 was considered statistically significant.

[0127] 2.2 Effects on catalase (CAT) activity

[0128] Wild-type AB strain zebrafish at 1 dpf were randomly selected and exposed to blue light to establish a zebrafish blue light eye damage model. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water (concentrations shown in Table 3-1), and a normal control group and a model control group were also set up. Each well contained 3 mL, and three biological replicates were performed. After treatment at 28℃ for 1 day, zebrafish samples were collected according to the CAT kit instructions. CAT activity was detected using a multi-mode microplate reader to analyze the CAT activity in zebrafish. The statistical analysis results were used to evaluate the effect of the samples on CAT activity. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software; p < 0.05 was considered statistically significant.

[0129] 3. Test Results

[0130] 3.1 Effect on MDA content

[0131] Blue light exposure causes oxidative damage to the eyes, including the oxidation of cellular lipids. Malondialdehyde (MDA) is one of the lipid oxidation products and can measure the severity of oxidative damage. Experimental results showed that after blue light exposure, the MDA content in the model control group was significantly increased compared to normal cells (p < 0.001), indicating that blue light exposure caused significant oxidative damage to ocular cellular lipids, and lipid oxidation led to a significant increase in MDA content. Compared with the model control group, monomeric taurine (250 μg / mL), monomeric coenzyme Q10 (40.0 μg / mL), and the combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) all significantly reduced the MDA content in the zebrafish eyes (p < 0.05 or p < 0.01, as shown in Table 3-1). Figure 7 As shown in the figure, the effects of taurine monomer (250 μg / mL) and the combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) on reducing MDA were more significant (p < 0.01 compared with the model control group). The treatment with the combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) had the lowest MDA content (1.88), which was lower than the average effect of using the same concentration of each ingredient alone (1.88 < 1.99). Therefore, this indicates that taurine and coenzyme Q10 have a synergistic effect at this ratio, and can achieve better results than using the same concentration of the monomer alone.

[0132] Table 3-1. Experimental results evaluating the impact of samples on malondialdehyde (MDA) content (n = 3)

[0133]

[0134] 3.2 Impact on CAT activity

[0135] Catalase (CAT) is an antioxidant enzyme widely distributed in organisms. It catalyzes the breakdown of hydrogen peroxide into water and oxygen, thereby protecting cells from oxidative damage caused by hydrogen peroxide. Experimental results show (as shown in Table 3-2 and...) Figure 8As shown in Table 3-2, CAT activity in the model control group was significantly lower than that in the normal control group, indicating successful model establishment. Compared with the model control group, both monomeric coenzyme Q10 (40.0 μg / mL) and the combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) significantly increased CAT activity in zebrafish eye cells (p < 0.05 or p < 0.01), indicating that both can play a significant protective role against blue light damage. Meanwhile, data showed that the CAT activity in zebrafish eye cells under the combined treatment (taurine and coenzyme Q10, concentrations of 250 and 40.0 μg / mL, respectively) was higher than the average effect of monomer treatment at the same concentration (Table 3-2, 0.497 > 0.466), indicating that taurine and coenzyme Q10 have a synergistic effect at this ratio, meaning that the combined treatment produces a greater effect than the individual treatment at the same concentration.

[0136] Table 3-2. Experimental results evaluating the effect of samples on catalase (CAT) activity (n = 3)

[0137]

[0138] Example 4: Evaluation of the impact on the expression of inflammation-related genes

[0139] 1. Testing materials

[0140] 1.1. Sample Preparation Information

[0141] Taurine was diluted with standard water, and coenzyme Q10 was dissolved in DMSO.

[0142] 1.2. Laboratory Animals

[0143] Zebrafish were raised in aquarium water at 28℃ (water quality: 200 mg of readily soluble sea salt added per 1 L of reverse osmosis water; conductivity 450~550 μS / cm; pH 6.5~8.5; hardness 50~100 mg / L CaCO3), bred and provided by Huante Biological Fish Farming Center. The laboratory animal use license number is: SYXK (Zhejiang) 2022-0004. The husbandry and management met the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0144] During the experiment, zebrafish were given different samples in the same manner as described in Example 1.

[0145] 1.3. Instruments, Consumables and Reagents

[0146] 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 Belamber Biotechnology Co., Ltd., China); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); Fully automatic sample grinder (JXFSTPRP-24L, Shanghai Jingxin Experimental Equipment Technology Department, China); Blue light analyzer (50W 450nm, China); High-speed refrigerated centrifuge (Heraeus Fresco17, Thermo Fisher, Germany); Fully automatic nucleic acid extractor (Auto-Pure32A, Hangzhou Aosheng Instrument Co., Ltd., China); Conventional PCR amplification instrument (T100, BIO-RAD, Singapore); Real-time PCR instrument (CFX). Connect, BIO-RAD, Singapore; UV-Vis 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-skirted 96-well plate (transparent) (HSP9601, Bio-rad, USA).

[0147] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Streptase 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 (Genomic De-generated) (Tiangen Biotech (Beijing) Co., Ltd., China); Pre-loaded Magnetic Bead Universal RNA Extraction Kit C (Foshan ONREW Biotechnology Co., Ltd., China).

[0148] 2. Detection Method

[0149] Wild-type AB strain zebrafish at 1 dpf were randomly selected and exposed to blue light to establish a zebrafish blue light eye damage model. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples and coenzyme Q10 (concentrations shown in Table 4-3) were administered in water, with a normal control group and a model control group also included. Each well contained 3 mL, and three biological replicates were performed. After treatment at 28℃ for 1 day, total RNA was extracted from each group of zebrafish using an automated nucleic acid extractor. The concentration and purity of total RNA were determined using a UV-Vis spectrophotometer. 2.00 μg of total RNA from the zebrafish sample was used to synthesize 20.0 μL of cDNA according to the cDNA first-strand synthesis kit instructions. The expression of β-actin and il-1β genes was detected by q-PCR. β-actin was used as an internal control for gene expression, and the relative RNA expression level of the il-1β gene was calculated. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and p < 0.05 indicated that the difference was statistically significant.

[0150] 3. Test Results

[0151] 3.1. RNA extraction results and primer sequence information

[0152] At the experimental endpoint, total RNA was extracted from zebrafish, and the RNA concentration and A260 / A280 ratio were measured using a UV-Vis spectrophotometer (Table 4-1). The A260 / A280 ratios were all between 1.8 and 2.2, indicating that the extracted total RNA from zebrafish was of good quality and could be used for subsequent q-PCR experiments. Primer sequences are shown in Table 4-2.

[0153] Table 4-1. Total RNA concentration and A260 / A280 ratio (n = 3)

[0154]

[0155] Table 4-2. Primer sequence information

[0156]

[0157] 3.2. Effects on the expression of inflammation-related genes

[0158] Interleukin-1β (IL-1β) is a key pro-inflammatory cytokine and one of the most important mediators of inflammation and host response to infection. It participates in various autoimmune inflammatory responses and multiple cellular activities, including cell proliferation, differentiation, and apoptosis. Overproduction of IL-1β is associated with pathophysiological changes occurring in different disease states, and IL-1β production is often increased in inflammatory infected cells.

[0159] The test results show (Table 4-3, Figure 9 After blue light irradiation, the relative expression level of the pro-inflammatory factor il-1β gene in the model control group was significantly increased, indicating that blue light irradiation caused a significant inflammatory response in the zebrafish eye cells. After treatment with certain concentrations of monomers and combinations, it was found that only the combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) significantly downregulated the relative expression level of the il-1β gene (p < 0.05), meaning that the combination could significantly improve the inflammatory response in the zebrafish eyes induced by blue light irradiation. This effect was also superior to the average effect of the monomers acting alone at the same concentration (0.578 < 0.647), indicating that taurine and coenzyme Q10 have a synergistic effect at this ratio, meaning that the combined use of the two can achieve effects that cannot be achieved by the monomers acting alone at the same concentration.

[0160] Table 4-3. Experimental results evaluating the effects of samples on the expression of inflammation-related genes (n = 3)

[0161]

[0162] Example 5: Evaluation of the impact on ocular histopathology

[0163] 1. Testing materials

[0164] 1.1. Sample Preparation Information

[0165] The solvents for lutein and coenzyme Q10 are both DMSO, and the solvent for taurine is standard dilution water.

[0166] 1.2. Laboratory Animals

[0167] Zebrafish were raised in aquarium water at 28℃ (water quality: 200 mg of readily soluble sea salt added per 1 L of reverse osmosis water; conductivity 450~550 μS / cm; pH 6.5~8.5; hardness 50~100 mg / L CaCO3), bred and provided by Huante Biological Fish Farming Center. The laboratory animal use license number is: SYXK (Zhejiang) 2022-0004. The husbandry and management met the requirements of international AAALAC certification (certification number: 001458), and the IACUC ethics review number is: IACUC-2023-6809-01.

[0168] During the experiment, zebrafish were given different samples in the same manner as described in Example 1.

[0169] 1.3. Instruments, Consumables and Reagents

[0170] 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 Beilanbo Biotechnology Co., Ltd., China); Ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China); Blue light analyzer (50W 450nm, China); Electronic thermostatic stainless steel water bath (HHS-2S, Shanghai Kanglu Instrument Equipment Co., Ltd., China); Microtome (KD2258, Jinhua Kedi Instrument Equipment Co., Ltd., China); Intelligent hot plate (400X280, Tianjin Laiyue Nag Laboratory Instrument Sales Co., Ltd., China); Biological microscope (CX31, OLYMPUS, Japan).

[0171] Dimethyl sulfoxide (DMSO, Shanghai Aladdin Biochemical Technology Co., Ltd., China); Streptase E (Shanghai Yuanye Biotechnology Co., Ltd., China); 4% tissue cell fixative (Beijing Solarbio Science & 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 resin (Beijing Solarbio Science & Technology Co., Ltd., China); High-efficiency sectioning paraffin (melting point 54-56℃, Shanghai Huayong Paraffin Co., Ltd., China); High-efficiency sectioning paraffin (melting point 62-64℃, Shanghai Huayong Paraffin Co., Ltd., China).

[0172] 2. Detection Method

[0173] Wild-type AB strain zebrafish with 1 day post-flop (dpf) were randomly selected and exposed to blue light to establish a zebrafish blue light-induced eye damage model. At 3 days post-flop (dpf), well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water (concentrations see [concentration details]). Figure 11 A normal control group and a model control group were set up, with a volume of 3 mL per well. After treatment at 28℃ for 1 day, zebrafish were fixed with 4% tissue cell fixative. Following a series of steps including dehydration, embedding, sectioning, and staining, the zebrafish underwent histopathological H&E staining analysis. The impact of the samples on the histopathology of zebrafish eyes was evaluated through ocular histopathological analysis.

[0174] 3. Test Results

[0175] Figure 10The image shows sections of zebrafish eye tissue analyzed and the names of each tissue. The experimental results show (…). Figure 11 In the normal control group, the zebrafish eye tissues were of normal size and shape, with clear and intact retinal structures and orderly arrangement of cells at all levels. Compared with the normal control group, the zebrafish in the model control group exposed to blue light showed a reduction in eye size (e.g., ...). Figure 11 As shown in ①, the retinal structure is disordered, lacks a hierarchical structure, and is arranged randomly (e.g., Figure 11 As shown in Figure ②), the outermost protective pigment epithelium becomes thinner and exhibits pigment diffusion, changing from black to light brown (as shown in Figure ②). Figure 11 As shown in ③), the cells in the cone and rod cell layers of the photoreceptors are arranged irregularly (e.g., Figure 11 As shown in ④), the outer nuclear layer and inner nuclear layer cells, which provide structural support, are arranged irregularly (e.g., Figure 11 As shown in Figure ⑤, all the above signs indicate that the zebrafish blue light eye damage model has been successfully established.

[0176] Compared with the model control group, the positive control group treated with lutein at a concentration of 1000 μg / mL showed significant recovery in zebrafish eye size. The retina exhibited a hierarchical and orderly arrangement, the pigment epithelium was significantly thickened without pigment diffusion, the cone and rod cell layers were regularly arranged, and the cell arrangement of the outer and inner nuclear layers was somewhat restored. The combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) also showed significant recovery in eye size, with the retina exhibiting a hierarchical and orderly arrangement, the pigment epithelium being significantly thickened without pigment diffusion, the cone and rod cell layers being regularly arranged, and the cell arrangement of the outer and inner nuclear layers being somewhat restored. Therefore, it can be concluded that treatment with lutein (1000 μg / mL) and the combination of taurine and coenzyme Q10 (250 + 40.0 μg / mL) resulted in some recovery of the eye tissue structure in zebrafish exposed to blue light, and these treatments have a mitigating and ameliorative effect on the damage to zebrafish eye tissue caused by blue light irradiation.

[0177] In summary, blue light, with its high energy, can cause oxidative damage to the zebrafish eyes at certain levels. This damage manifests as the abnormal accumulation of reactive oxygen species (ROS) and a decrease in the activity of ocular antioxidant enzymes (such as catalase, CAT), leading to increased lipid oxidation. Consequently, lipid oxidation-related products such as malondialdehyde (MDA) also increase. Oxidative damage results in the upregulation of the relative expression of pro-inflammatory factors such as IL-1β, inducing an inflammatory response and ultimately leading to apoptosis of zebrafish ocular cells and even abnormal changes in tissue structure. This study found that certain concentrations of taurine monomers, coenzyme Q10 monomers, and combinations of taurine and coenzyme Q10 can alleviate damage caused by blue light irradiation. Specifically, this is manifested in reducing the degree of ocular cell apoptosis, scavenging reactive oxygen species in the eye, reducing the content of lipid peroxidation products (MDA), increasing the activity of antioxidant enzymes (CAT), reducing the relative expression of the pro-inflammatory factor IL-1β gene il-1β, inhibiting inflammation, and improving the structure and condition of damaged ocular tissues. Comparative analysis revealed that certain ratios of taurine + coenzyme Q10 combinations ((0.17~7.14):1) can all reduce ocular cell apoptosis caused by blue light irradiation to a certain extent (even though the effective amounts of both in the combination are far less than the effective amounts of the monomers), and the effect of the combination is superior to the average effect of using the same concentration of monomers alone. This means that these combinations have a synergistic effect, and their combined use can achieve effects that cannot be achieved by using the monomers alone. Further research revealed that the lowest levels of ocular cell apoptosis were observed when the concentrations of taurine and coenzyme Q10 were 80.0 ± 40.0, 250 ± 35.0, and 250 ± 40.0 µg / mL (in a ratio of (2.00–7.14):1). This indicates that these concentration combinations resulted in less blue light damage to the zebrafish eyes, demonstrating the superior efficacy of the combination. Further comparison revealed that the combination at a concentration of 250 ± 40.0 µg / mL (in a ratio of 6.25:1) exhibited the best anti-apoptotic effect on ocular cells. This effect was significantly better than that of coenzyme Q10 alone at a concentration of 40.0 µg / mL (p < 0.05), and also significantly better than the average effect of either ingredient alone (p < 0.05). Therefore, it can be concluded that the synergistic effect of the two at this ratio was the most significant. Using this combination as an example, further studies confirmed that the combination of taurine and coenzyme Q10 can effectively scavenge reactive oxygen species generated in the eyes of zebrafish under blue light irradiation, reduce the content of lipid peroxidation 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 inflammation, and improve the structure and condition of damaged eye tissues. Moreover, the above effects of the combination are all superior to the average effect of the monomeric components used alone at the same concentration, thus exhibiting a synergistic effect.

[0178] Industrial availability

[0179] The compositions provided by this invention can be mass-produced and used in industry.

Claims

1. The use of a functional composition in mitigating blue light damage, characterized in that, The functional composition comprises the following components: Taurine and Coenzyme Q10; Furthermore, in the functional composition, the effective amount ratio of taurine to coenzyme Q10 is (6.0~6.5):1, the effective amount of taurine is greater than or equal to 6.71 μg / mL, and the effective amount of coenzyme Q10 is greater than or equal to 30 μg / mL. The reduction of blue light damage involves reducing the accumulation of reactive oxygen species in the eyes, reducing the content of lipid oxidation products in the eyes, and increasing the activity of antioxidant enzymes in the eyes.

2. The use according to claim 1, characterized in that, The reduction of ocular lipid oxidation product content includes reducing ocular malondialdehyde content.

3. The use according to claim 1, characterized in that, The enhancement of ocular antioxidant enzyme activity includes enhancing ocular catalase activity.

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