Aging eyeball model and construction method and application thereof

Abstract

The invention relates to the technical field of eyeball model construction, in particular to an aging eyeball model and a construction method and application thereof. The invention provides an aging eyeball model construction method, which comprises the following steps that an N-sub-retinyl-N-retinyl-ethanolamine solution is prepared, and the concentration of the N-sub-retinyl-N-retinyl-ethanolamine in the N-sub-retinyl-N-retinyl-ethanolamine solution is 20 to 80 mol / L, and the concentration of the N-sub-retinyl-N-retinyl-ethanolamine in the N-sub-retinyl-N-retinyl-ethanolamine solution is 20 to 80 mol / L; an N-sub-retinyl-N-retinyl-ethanolamine solution is added into animal drinking water, the animal drinking water is irradiated with blue light, the aging eyeball model is obtained, and the adding amount of the N-sub-retinyl-N-retinyl-ethanolamine solution in the drinking water is 1-50% by weight. According to the method, the senescence eyeball model or the age-related macular degeneration animal model can be quickly constructed, and the construction period of the senescence eyeball model or the age-related macular degeneration animal model is shortened.

Description

Technical Field

[0001] This invention relates to the field of eyeball model construction technology, specifically to an aging eyeball model and its construction method. Background Technology

[0002] The eyes are exposed to the external environment and are easily affected by adverse factors such as strong light, high pressure, and bacteria and viruses. This forces the eyes to constantly defend against these factors to prevent more serious eye diseases. However, with age, this resistance weakens. Simultaneously, eye function declines over time, and the ability to receive, store, and analyze visual information diminishes. Some eye diseases become more prevalent due to aging, such as age-related macular degeneration (AMD), retinopathy, glaucoma, cataracts, vision loss, and eye strain. Currently, most research uses mouse eyes as models to establish corresponding ophthalmic disease models for disease mechanism analysis and drug screening. However, experimental modeling is difficult, time-consuming, and involves complex evaluation indicators. Especially when rapidly screening drugs to improve or combat aging, the inaccurate aging degree of the eye model and the long construction time hinder research and development. Summary of the Invention

[0003] Therefore, the technical problem to be solved by the present invention is to overcome the defects of inaccurate construction of the degree of aging and unstable shape of the eyeball model in the prior art, thereby providing an aging eyeball model and its construction method and application.

[0004] To address the aforementioned technical problems, this invention provides a method for constructing an aging eyeball model, comprising the following steps.

[0005] Prepare an N-sub-retinyl-N-retinyl-ethanolamine solution, wherein the concentration of N-sub-retinyl-N-retinyl-ethanolamine in the N-sub-retinyl-N-retinyl-ethanolamine solution is 20-80 mol / L;

[0006] An aging eye model was obtained by adding N-sub-retinyl-N-retinyl-ethanolamine solution to animal drinking water and irradiating the animals with blue light. The amount of N-sub-retinyl-N-retinyl-ethanolamine solution added to the drinking water was 1 wt‰ to 50 wt‰.

[0007] Furthermore, the solvent in the N-sub-retinyl-N-retinyl-ethanolamine solution includes water.

[0008] Furthermore, the duration of blue light irradiation is 5-14 days.

[0009] Furthermore, the intensity of the blue light is 1000-5000 lux.

[0010] In some alternative embodiments, the amount of N-retinyl-N-retinyl-ethanolamine added to the drinking water is 1 wt‰ to 18 wt‰.

[0011] Furthermore, the animal is at least one of zebrafish, rat, rabbit, or mouse.

[0012] In some alternative embodiments, the animal is a rat, the blue light intensity is 1000-2300 lux, and the blue light exposure time is 7-10 days.

[0013] In some optional embodiments, the method for constructing an aging eye model further includes the step of fixing the aging eye model with a fixative.

[0014] In some optional embodiments, the method for constructing an aging eye model further includes a step of detecting the degree of aging of the aging eye model, wherein the target substances for detection include superoxide dismutase and / or malondialdehyde.

[0015] In some alternative implementations, the aging eye model includes an animal model of age-related macular degeneration.

[0016] The present invention also provides an aging eye model, which is constructed according to the above-described method for constructing an aging eye model.

[0017] The present invention also provides the application of the above-mentioned aging eye model in screening and preparing drugs, health foods, health care products or products that improve or resist eye aging.

[0018] The technical solution of this invention has the following advantages:

[0019] This invention provides a method for constructing an aging eye model, comprising the following steps: preparing an N-sub-retinyl-N-retinyl-ethanolamine solution, wherein the concentration of N-sub-retinyl-N-retinyl-ethanolamine in the N-sub-retinyl-N-retinyl-ethanolamine solution is 20-80 mol / L; adding the N-sub-retinyl-N-retinyl-ethanolamine solution to animal drinking water; and irradiating the animals with blue light after drinking the water to obtain an aging eye model, wherein the amount of N-sub-retinyl-N-retinyl-ethanolamine added to the drinking water is 1 wt‰ to 50 wt‰. This invention enables rapid construction of an aging eye model by injecting the N-sub-retinyl-N-retinyl-ethanolamine solution into the fundus or by adding the N-sub-retinyl-N-retinyl-ethanolamine solution to animal drinking water, followed by blue light irradiation, thus shortening the construction cycle of the aging eye model.

[0020] Furthermore, the method for constructing an aging eye model provided by this invention can quickly construct an animal model of age-related macular degeneration, shortening the construction cycle of the animal model of age-related macular degeneration.

[0021] Meanwhile, the method for constructing an aging eye model provided by this invention can control the degree of aging in the eye model by adjusting the amount of N-sub-retinyl-N-retinyl-ethanolamine added, the distance between the blue light and the animal, and the irradiation time. This allows for accurate screening of ophthalmic products such as drugs and health foods, as well as the accurate acquisition of ophthalmic products with different degrees of aging. The aging eye model obtained by controlling the amount of N-sub-retinyl-N-retinyl-ethanolamine added to drinking water between 1wt‰ and 50wt‰ exhibits a more significant atrophy effect and a superior aging effect. Attached Figure Description

[0022] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0023] Figure 1 These are electroretinograms of rats with an aging eyeball model constructed in Examples 1-8 of Experiment 1 of this invention;

[0024] Figure 2 These are electroretinograms of the aging eyeball model rats constructed in Examples 9-15 of Experimental Example 1 of this invention;

[0025] Figure 3 It is the electroretinogram of the aging eyeball model rat constructed in Comparative Examples 1-4 in Experimental Example 1 of this invention;

[0026] Figure 4 This is a cataract image of the eyeball of the rat model of aging eyeball constructed in Comparative Example 1 of this invention;

[0027] Figure 5 These are the visual black-and-white box results of the aging eyeball model rat constructed in Examples 1-5 of Experiment 2 of this invention;

[0028] Figure 6 These are the visual black-and-white box results of the aging eyeball model rat constructed in Examples 6-10 of Experiment 2 of this invention;

[0029] Figure 7 These are the visual black-and-white box results of the aging eyeball model rat constructed in Examples 11-15 of Experimental Example 2 of this invention;

[0030] Figure 8 This is a black-and-white box result of the visual model rats with aging eyeballs constructed in Comparative Examples 1-4 of Experimental Example 2 of this invention;

[0031] Figure 9This is an image showing the HE staining results of the eyes of the rats used in the aging eyeball model constructed in Examples 1-8 of Experiment 3 of this invention;

[0032] Figure 10 This is an image showing the HE staining results of the eyes of the rats used in the aging eyeball model constructed in Examples 9-15 of Experimental Example 3 of this invention;

[0033] Figure 11 This is an HE staining result of the eyes of rats in the aging eyeball model constructed in Comparative Examples 1-4 of Experimental Example 3 of this invention;

[0034] Figure 12 This is a diagram showing the ROS staining results of the eyes of the rats that formed the aging eyeball model in Examples 1-5 of Experiment 3 of this invention;

[0035] Figure 13 This is a diagram showing the ROS staining results of the eyes of rats in the aging eyeball model constructed in Examples 6-10 of Experiment 3 of this invention;

[0036] Figure 14 This is a diagram showing the ROS staining results of the eyes of the rats that formed the aging eyeball model in Examples 11-15 of Experiment 3 of this invention;

[0037] Figure 15 This is a diagram showing the ROS staining results of the eyes of rats that formed the aging eyeball model in Comparative Examples 1-4 of Experimental Example 3 of this invention. Detailed Implementation

[0038] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0039] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0040] Example 1

[0041] This embodiment provides a method for constructing an aging eyeball model, with the specific steps and parameters as follows:

[0042] A 35 mol / L N-sub-retinyl-N-retinyl-ethanolamine (A2E) solution was prepared using deionized water;

[0043] Add 18wt‰ A2E solution to the drinking water of rats (weighing 18±2g). The rats were housed in an SPF-grade environment and simultaneously irradiated with blue light using an animal photodamage device (patent publication number CN221429878U). The light intensity was 5000 lux and the irradiation lasted for 5 days. After the irradiation ended, the rats were sacrificed, the eyeballs were removed, and the eyeballs were fixed with 10Vol% formalin fixative to obtain an aging eyeball model.

[0044] Example 2

[0045] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the amount of A2E solution added to the drinking water is 1 wt‰.

[0046] Example 3

[0047] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the amount of A2E solution added to the drinking water is 25wt‰.

[0048] Example 4

[0049] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the amount of A2E solution added to the drinking water is 50 wt‰.

[0050] Example 5

[0051] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the irradiation time of the blue light tube is 7 days.

[0052] Example 6

[0053] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the irradiation time of the blue light tube is 10 days.

[0054] Example 7

[0055] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the irradiation time of the blue light tube is 14 days.

[0056] Example 8

[0057] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the light intensity of the blue light tube is 1000 lux.

[0058] Example 9

[0059] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the light intensity of the blue light tube is 2300 lux.

[0060] Example 10

[0061] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the light intensity of the blue light tube is 3000 lux.

[0062] Example 11

[0063] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that the light intensity of the blue light tube is 4000 lux.

[0064] Example 12

[0065] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that a 20 mol / L N-sub-retinyl-N-retinyl-ethanolamine (A2E) solution is prepared using deionized water.

[0066] Example 13

[0067] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1, except that an 80 mol / L N-sub-retinyl-N-retinyl-ethanolamine (A2E) solution is prepared using deionized water.

[0068] Example 14

[0069] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1. The difference is that an 80 mol / L N-sub-retinyl-N-retinyl-ethanolamine (A2E) solution is prepared using deionized water, and the amount of A2E solution added to drinking water is 50 wt‰.

[0070] Example 15

[0071] This embodiment provides a method for constructing an aging eyeball model. The specific steps and parameters are the same as in Embodiment 1. The difference is that a 20 mol / L N-sub-retinyl-N-retinyl-ethanolamine (A2E) solution is prepared using deionized water, and the amount of A2E solution added to drinking water is 1 wt‰.

[0072] Comparative Example 1

[0073] This comparative example provides a method for constructing an aging eyeball model. The specific steps and parameters are as follows:

[0074] A solution of N-sub-retinyl-N-retinyl-ethanolamine (A2E) with a concentration of 80 mol / L was prepared using deionized water;

[0075] Add 52wt‰ A2E solution to the drinking water of rats and house them in an SPF-grade environment. Simultaneously, irradiate the rats with blue light using an animal photodamage device (patent publication number CN221429878U). The light intensity is 5000 lux and the irradiation lasts for 5 days. After the irradiation ends, fix the eyeballs with 10Vol% formalin fixative to obtain an aging eyeball model.

[0076] Comparative Example 2

[0077] This comparative example provides a method for constructing an aging eyeball model. The specific steps and parameters are as follows:

[0078] A 20 mol / L N-retinyl-N-retinyl-ethanolamine (A2E) solution was prepared using deionized water.

[0079] Add 0.5 wt‰ A2E solution to the drinking water of rats and house them in an SPF-grade environment. Simultaneously, irradiate the rats with blue light using an animal photodamage device (patent publication number CN221429878U). The light intensity is 5000 lux and the irradiation lasts for 5 days. After the irradiation ends, fix the eyeballs with 10 vol% formalin fixative to obtain an aging eyeball model.

[0080] Comparative Example 3

[0081] This comparative example provides a method for constructing an aging eyeball model. The specific steps and parameters are as follows:

[0082] Rats were housed in an SPF-grade environment and subjected to blue light irradiation using an animal photodamage device (patent publication number CN221429878U). The light intensity was 5000 lux, and the irradiation lasted for 5 days. After the irradiation ended, the eyeballs were fixed with 10 vol% formalin fixative to obtain an aging eyeball model.

[0083] Comparative Example 4

[0084] This comparative example provides a method for constructing an aging eyeball model. The specific steps and parameters are as follows:

[0085] A 35 mol / L N-sub-retinyl-N-retinyl-ethanolamine (A2E) solution was prepared using deionized water;

[0086] Add 18wt‰ A2E solution to the drinking water of rats, house the rats in an SPF-grade environment, expose them to natural light (400 lux) for 5 days, and fix the eyeballs with 10 vol% formalin fixative to obtain an aging eyeball model.

[0087] Experiment 1 (Visual Electrophysiology)

[0088] In Examples 1-15 and Comparative Examples 1-4, after the rats were acclimatized to a dark environment for more than one day following the end of blue light or natural light exposure, the animals in each group were anesthetized to begin the experiment. Mydriasis was achieved using compound tropicamide eye drops. Electroretinography (ERG) was then performed in darkness (a weak red light could be used). One drop of levofloxacin hydrochloride ophthalmic gel was instilled into each eye of the rat, followed by placing the recording electrode on the cornea, and placing the ground electrode and reference electrode under the skin near the base of the head and tail, respectively. A-waves and b-waves were measured in each group of animals using a visual electrophysiological instrument.

[0089] The amplitudes of the a and b waves in ERG reflect the receptor function and electrical activity of the retina after photoreceptor stimulation, respectively. Figure 1-3 The results showed that the amplitudes of the a-wave (reflecting photoreceptor function) and b-wave (reflecting bipolar cell activity) in Examples 1-15 were significantly decreased, indicating that retinal function decline leads to ocular aging; while the amplitude decrease in Comparative Examples 2-4 was not significant. Although the amplitudes of the a-wave and b-wave in Comparative Example 1 were significantly decreased, ... Figure 4 The excessive A2E solution shown resulted in severe damage to the animal's eyeballs, including cataracts, near-total vision loss, and self-mutilation, rendering the animal unsuitable as a model for further research. In conclusion, this demonstrates that the ocular aging model constructed in this invention is more effective.

[0090] Experiment Example 2 (Visual Black and White Box)

[0091] (1) In all behavioral experiments, the animals should be frequently petted in the early stages to familiarize them with the operator's scent, thereby reducing errors caused by human interference during the behavioral experiment. The visual black-and-white box behavioral instrument should be placed in a separate, light-proof and soundproof room. During the test, excessive noise and strong odors should be avoided to prevent the rats from experiencing anxiety and affecting the experiment. Before placing each rat into the test box, both boxes should be cleaned with 75% ethanol to prevent the scent of the previous rat from affecting the rat's movement trajectory during the test. After wiping the boxes and waiting for the alcohol odor to evaporate, the animal should be petted while being transferred to one side of the box, allowing each animal at least 15 minutes to adapt before recording the animal's movement trajectory.

[0092] (2) After the blue light irradiation ended, all experimental rats in Examples 1-15 and Comparative Examples 1-4 were subjected to dark treatment overnight before the experiment. The lighting intensity in the white box was set to 200-1000 lux, referring to the article (Qin Huan, Gene Therapy and Behavioral Analysis of Retinal Degeneration Caused by PDE6B Mutation, Wuhan University of Science and Technology, 2023). No light source was provided in the black box. After the animals had adapted, the test was started. The infrared camera function built into the device was used to record the movement trajectory of the rats as they moved freely between the two boxes for 10 minutes.

[0093] like Figure 5-8 The results showed that in the visual black-and-white box experiment, the rats in Examples 1-15 spent a significantly longer time in the white box and exhibited disordered movement trajectories, suggesting severe impairment of light signal transduction and nerve transmission functions leading to ocular aging. In contrast, the rats in Comparative Examples 2-4 showed no significant change in their time spent in the white box. Although the rats in Comparative Example 1 also spent a significantly longer time in the white box, the excessive A2E solution caused severe damage to their eyeballs, resulting in cataracts, near-loss of vision, and self-harm, making them unsuitable as animal models for further research. In summary, this invention demonstrates that the ocular aging model constructed in this invention is more effective.

[0094] Experimental Example 3

[0095] The aging eye models obtained in Examples 1-15 and Comparative Examples 1-4 were immediately removed from the eyeballs. One eyeball was placed in a tissue fixation solution, and the other eyeball was retinally detached and placed in a grinding tube containing PBS and grinding beads. The mixture was then homogenized in a homogenizer, and the supernatant was collected for testing.

[0096] Three days later, HE staining and ROS staining were performed to examine the ocular photoaging of each model. HE staining results provide basic morphological evidence for assessing tissue structure and pathological changes, and the degree of damage can be quantitatively determined by comparing the thickness of the outer nuclear layer (ONL) of the retina in each group. ROS directly assesses key pathogenic factors and oxidative stress levels in aging eyes; the higher the expression level, the greater the degree of aging.

[0097] The HE staining steps are as follows:

[0098] One eyeball from each of the experimental mice was harvested and the following steps were performed:

[0099] (1) Dehydration and embedding: After the tissue sample is loaded into a standardized dehydration box, it is subjected to a gradient ethanol replacement program through a fully automated tissue dehydration system, and then the tissue embedding preparation is completed.

[0100] (2) Sectioning: Cut 4μm tissue sections with a microtome, flatten them in water, remove them with a glass slide, and place them on a curing machine at 60℃ for 1 hour.

[0101] (4) Staining: After baking the slides, put the white slides of each tissue into the fully automatic staining machine and click the program to start the staining program.

[0102] (5) Mounting and observation: After staining, mount the slides immediately with neutral resin, then transfer them to a ventilated place overnight, and observe them under a microscope the next day.

[0103] ROS staining steps:

[0104] The other eyeball of each of the experimental mice was removed and the following steps were performed:

[0105] Remove the animal eyeballs from the -80℃ freezer, adjust the microtome to 5μm for sectioning, and place them on frozen microscope slides.

[0106] The prepared frozen sections were placed together with dihydroethidium (DHE) probe working solution in a 37°C incubator for a light-protected reaction for 30 minutes. After the cell nuclei were fixed, the sections were sealed with anti-fluorescence quenching slides containing DAPI for observation.

[0107] Replenish experimental supplies:

[0108] Table 1 Main Reagents and Consumables

[0109]

[0110] Table 2 Main Instruments and Consumables

[0111]

[0112] See results Figure 9-11 HE staining results showed that the outer nuclear layer of the retina in Examples 1-15 all exhibited varying degrees of atrophy; while in Comparative Examples 2-4, the change in the outer nuclear layer thickness was not significant. Although the outer nuclear layer of the retina in Comparative Example 1 also showed atrophy, the excessive A2E solution caused severe damage to the animal's eyeball, resulting in cataracts, near-loss of vision, and self-mutilation, making it unsuitable as an animal model for other studies. In summary, this invention demonstrates that the aging eye model constructed can exhibit obvious and appropriate aging eye characteristics, with severe functional degeneration of the retinal ONL and photoreceptor layer.

[0113] ROS is a peroxide that accumulates in large quantities in aging areas, as shown in the staining results. Figure 12-15The results show that Examples 1-15 all exhibited varying degrees of excessive ROS accumulation, while Comparative Examples 2-4 showed less significant ROS accumulation. Although Comparative Example 1 also showed substantial ROS accumulation, the excessive A2E solution resulted in severe damage to the animal's eyeball, leading to cataracts, near-loss of vision, and self-harm, rendering it unsuitable as an animal model for further research. In conclusion, this demonstrates that the aging eyeball model constructed in this invention exhibits better aging response.

[0114] Experiment Example 4

[0115] Retinas from rats in Examples 1-15 and Comparative Examples 1-4 were placed in homogenization tubes, and an appropriate volume of PBS was added according to the recommended dilution concentration in the manufacturer's instructions. The homogenates were then sonicated at -10°C to obtain tissue homogenates for each group. Subsequently, the supernatant from each tissue homogenate was centrifuged at 3000 rpm for 10 min using a low-temperature centrifuge. The SOD and MDA contents were then determined according to the kit instructions. Before performing the tests on each indicator, the protein concentration of the samples was determined using a BCA kit for calculation. The results are shown in Table 3.

[0116] SOD content detection method: The total superoxide dismutase (T-SOD) assay kit (WST-1 method) from Nanjing Jiancheng was used. The detection was performed according to the instructions. The SOD results were obtained.

[0117] MDA content detection method: The Nanjing Jiancheng Malondialdehyde (MDA) assay kit (TBA method) was used to perform the test according to the instructions. The MDA results were obtained.

[0118] Table 3. Content of SOD and MDA in the retina of experimental rats

[0119]

[0120]

[0121] SOD (superoxide dismutase) helps scavenge free radicals and reduce and eliminate oxidative damage; lower SOD levels indicate a higher degree of aging. MDA (malondialdehyde), a lipid peroxidation product, also indicates a higher degree of aging. Table 3 shows that compared to Comparative Examples 2-4, Examples 1-15 showed lower SOD levels and higher MDA levels in the rat retinas. Although Comparative Example 1 had low SOD and high MDA levels, the excessive A2E solution caused severe eye damage, cataracts, near-loss of vision, and self-harm, making it unsuitable as an animal model for further research. In conclusion, the aging eye model constructed in this invention demonstrates better aging response.

[0122] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A method for constructing an aging eyeball model, characterized in that, Includes the following steps, Prepare an N-sub-retinyl-N-retinyl-ethanolamine solution, wherein the concentration of N-sub-retinyl-N-retinyl-ethanolamine in the N-sub-retinyl-N-retinyl-ethanolamine solution is 20-80 mol / L; An aging eye model was obtained by adding N-sub-retinyl-N-retinyl-ethanolamine solution to animal drinking water and irradiating the animals with blue light. The amount of N-sub-retinyl-N-retinyl-ethanolamine solution added to the drinking water was 1 wt‰ to 50 wt‰.

2. The method for constructing an aging eyeball model according to claim 1, characterized in that, The solvent in the N-sub-retinyl-N-retinyl-ethanolamine solution includes water.

3. The method for constructing an aging eyeball model according to claim 1 or 2, characterized in that, The duration of blue light irradiation is 5-14 days.

4. The method for constructing an aging eyeball model according to any one of claims 1-3, characterized in that, The intensity of the blue light is 1000-5000 lux.

5. The method for constructing an aging eyeball model according to any one of claims 1-4, characterized in that, The amount of N-retinyl-N-retinyl-ethanolamine added to the drinking water is 1 wt‰ to 18 wt‰.

6. The method for constructing an aging eyeball model according to any one of claims 1-5, characterized in that, The animal is at least one of zebrafish, rat, rabbit or mouse.

7. The method for constructing an aging eyeball model according to any one of claims 1-6, characterized in that, The animal in question is a rat, the intensity of the blue light is 1000-2300 lux, and the duration of blue light exposure is 7-10 days. The method for constructing an aging eye model also includes the step of fixing the aging eye model with a fixative.

8. The method for constructing an aging eyeball model according to any one of claims 1-7, characterized in that, The method for constructing an aging eye model further includes a step of detecting the degree of aging in the aging eye model, wherein the target substances for detection include superoxide dismutase and / or malondialdehyde; and / or, The aging eye model includes an animal model of age-related macular degeneration.

9. An aging eyeball model, characterized in that, The aging eye model is constructed by the method for constructing an aging eye model as described in any one of claims 1-8.

10. The application of the aging eye model according to claim 9 in screening and preparing drugs, health foods, health products or products for improving or resisting eye aging.

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