Method for constructing visual fatigue animal model
By constructing an animal model of visual fatigue through controlling light intensity and dark adaptation, the problems of excessively long treatment cycles and irreversible damage were solved, thus achieving the reversibility of visual fatigue and the accuracy of experimental results.
Patent Information
- Application Number
- CN202410138016.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-02-01
- Publication Date
- 2026-02-06
- Estimated Expiration
- 2044-02-01
AI Technical Summary
Existing animal models of visual fatigue have excessively long cycles, or the experimental animals suffer irreversible visual damage.
By controlling the light intensity of the experimental light source within the range of 1000 lux to 5000 lux, experimental animals were subjected to light exposure experiments. After the light cycle ended, dark adaptation was performed, and combined with appropriate humidity and physiological saline eye drops, an animal model of visual fatigue was constructed.
While ensuring the reversibility of visual impairment, it avoids the increase in blood lipid levels caused by excessively long experimental periods, ensuring the accuracy and reliability of experimental results, and preventing visual fatigue recovery.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of light damage, and particularly relates to a method for constructing a visual fatigue animal model. BACKGROUND
[0002] The eye is an important organ for human beings to acquire knowledge and memory. When the eye works intensively, the eye is prone to visual fatigue, such as blurred vision, eye acid swelling, dryness, tearing and other eye discomfort, due to long-time accommodation. Visual fatigue not only affects the efficiency of work and study, but also easily affects physical and mental health for a long time.
[0003] At present, experimental animals are often used for visual fatigue experiments. However, the existing visual fatigue animal model has a long cycle, or the experimental animals have irreversible visual damage. SUMMARY
[0004] Therefore, the present application provides a method for constructing a visual fatigue animal model to solve the problems of long cycle of the visual fatigue animal model or irreversible visual damage of the experimental animals.
[0005] In a first aspect, the present application provides a method for constructing a visual fatigue animal model, comprising:
[0006] placing the experimental animals in a light-free condition for a first dark adaptation;
[0007] randomly dividing the experimental animals into a normal group and an experimental group, respectively, performing a light exposure experiment on the normal group and the experimental group, feeding the normal group in an environment simulating daily light exposure, feeding the experimental group in an animal light damage device, and placing the animal light damage device in an environment with a light intensity of 1000 lux to 5000 lux of the experimental light source;
[0008] performing an experiment on the experimental animals for a light cycle t;
[0009] detecting physical indicators of the experimental animals.
[0010] Beneficial effects: by controlling the light intensity of the experimental light source, reversible visual damage of the experimental animals is ensured, and the experimental cycle is prevented from being too long, the blood lipid indicators of the experimental animals are prevented from being improved, and the model construction is prevented from being affected and the experimental results are prevented from being disturbed.
[0011] In an optional embodiment, after the light cycle ends, the experimental animals are first placed in a light-free condition for a second dark adaptation, and then the physical indicators of the experimental animals are detected.
[0012] Beneficial effect: the second dark adaptation after the light cycle helps to further consolidate visual fatigue, and also helps to avoid animal stress caused by light experiments affecting the authenticity of the experiment.
[0013] In an alternative embodiment, the second dark adaptation period is 5 to 15 days.
[0014] Beneficial effect: while ensuring that dark adaptation consolidates visual fatigue, it prevents damage from starting to recover due to prolonged dark adaptation.
[0015] In an alternative embodiment, during the second dark adaptation, the experimental animals are only provided with maintenance feed to maintain life.
[0016] Beneficial effect: providing the experimental animals with only maintenance feed, the experimental animals only receive nutrients for life, which helps to further consolidate visual fatigue and prevent experimental animals from recovering from visual fatigue due to excessive nutrition.
[0017] In an alternative embodiment, during the light cycle, 10% to 20% physiological saline is dropped into the eyes of the experimental animals before light exposure.
[0018] Beneficial effect: using 10% to 20% physiological saline to drop into the eyes of the experimental animals increases the osmotic pressure of the ocular surface, making it more susceptible to light exposure and causing fatigue.
[0019] In an alternative embodiment, the light cycle t is 20 to 40 days.
[0020] In an alternative embodiment, the light cycle is a 12-hour / 12-hour light-dark cycle.
[0021] In an alternative embodiment, the experimental group is placed in an experimental environment with a humidity of 20% to 30%.
[0022] Beneficial effect: placing the experimental group in a relatively dry environment can accelerate the evaporation of water from the ocular surface, leading to frequent eye dryness and more easily causing fatigue, further inducing the severity of visual fatigue.
[0023] In an alternative embodiment, the experimental animals are rats, and the rats are 6 to 20 weeks old.
[0024] Beneficial effect: selecting rats aged 6 to 20 weeks, the size of the eyeball is suitable for experimental sampling, and also meets the required amount for biochemical index detection, which is more conducive to experimental operation.
[0025] In an alternative embodiment, the experimental light source is ultraviolet light, blue light or green light. DETAILED DESCRIPTION
[0026] To make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.
[0027] According to an embodiment of the present application, in one aspect, a method for constructing a visual fatigue animal model is provided, comprising: placing experimental animals in a lightless condition for a first dark adaptation; randomly dividing the experimental animals into a normal group and an experimental group, and respectively performing light exposure experiments on the normal group and the experimental group, feeding the normal group in an environment simulating daily light exposure, and feeding the experimental group in an animal light damage device, and placing the animal light damage device in an environment with a light intensity of 1000 lux to 5000 lux of an experimental light source; performing an experiment on the experimental animals for a light cycle t; and detecting physical indicators of the experimental animals.
[0028] The method for constructing a visual fatigue animal model in the embodiment can control the light intensity of the experimental light source, ensure reversible visual damage of the experimental animals, prevent the experimental animals from having improved blood lipid indicators due to a long experimental period, and interfere with the experimental results.
[0029] It should be noted that the light intensity is a key factor for modeling. A light source with a light intensity lower than 1000 lux is not enough to cause retinal damage of the animals, which leads to a prolonged modeling period, a prolonged growth period of the experimental animals, high feeding costs of the animals, and improved blood lipid indicators of the animals (because the experimental animals are fed for a long time in cages without exercise, which inevitably causes an increase in blood lipid of the animals), which is not conducive to model construction. A light source with a light intensity higher than 5000 lux can cause irreversible light damage, which is inconsistent with actual needs, and therefore cannot exceed the range.
[0030] It should be noted that the experimental animals are placed in a lightless condition for dark adaptation to prevent the experimental animals from having repulsion and stress to the environment, which is not conducive to feeding, gavage and the like in the experimental process.
[0031] It should be noted that the experimental animals are killed at the same time to ensure consistency of the experimental detection period.
[0032] It should be noted that the drinking water and feed of all the experimental animals are consistent.
[0033] In one embodiment, after the end of the light period, the experimental animals are first placed in a lightless condition for a second dark adaptation, and then the physical indicators of the experimental animals are detected.
[0034] It is worth noting that the second dark adaptation after the end of the light period helps to further consolidate visual fatigue, and also helps to avoid excessive stress of the animals caused by the light experiment affecting the authenticity of the experimental detection.
[0035] In one embodiment, the period of the second dark adaptation is 5 to 15 days.
[0036] It should be noted that if the experimental animals are in the second dark adaptation for too long, the visual fatigue damage will start to recover slowly. That is, after the adverse conditions are removed, the visual fatigue damage will first be aggravated for a period of time, and then start to recover slowly. Therefore, the period of dark damage is very important.
[0037] It is worth noting that while ensuring that dark adaptation consolidates visual fatigue, it prevents damage from starting to recover due to too long dark adaptation time.
[0038] In one embodiment, during the second dark adaptation, the experimental animals are only provided with maintenance feed to maintain life.
[0039] It is worth noting that the experimental animals are only provided with maintenance feed, and the experimental animals only obtain nutrients for maintaining life, which is conducive to further consolidating visual fatigue and preventing the experimental animals from recovering from visual fatigue due to excessive nutrition.
[0040] In one embodiment, during the light period, 10% to 20% physiological saline is dropped in the eyes of the experimental animals before light exposure every day.
[0041] It is worth noting that using 10% to 20% physiological saline to drop in the eyes of the experimental animals increases the osmotic pressure of the ocular surface, making it more susceptible to light exposure and causing fatigue.
[0042] In one embodiment, the light period t is 20 to 40 days.
[0043] It should be noted that under the condition of a certain light intensity, according to the selection of the light period, a mild, moderate, and high visual fatigue animal model can be constructed.
[0044] In one embodiment, the light mode of the light period is a 12-hour / 12-hour light-dark cycle.
[0045] It should be noted that the light mode of 12 hours / 12 hours light-dark cycle conforms to the regularity of the animal's work and rest, preventing the animal's work and rest from being disturbed and affecting the experimental results.
[0046] In one embodiment, the experimental group is placed in an experimental environment with humidity of 20% to 30%.
[0047] It should be noted that in a normal feeding environment, the humidity is generally 40% to 70%, and the appropriate environmental humidity is beneficial to provide a suitable moisture environment for the eyes, and a dry environment can easily accelerate the evaporation of moisture on the ocular surface, leading to frequent ocular dryness, and cooperating with the light of the experimental light source, it is more likely to further induce the severity of visual fatigue.
[0048] It should be noted that placing the experimental group in a relatively dry environment can easily accelerate the evaporation of moisture on the ocular surface, leading to frequent ocular dryness, and more likely to produce fatigue, further inducing the severity of visual fatigue.
[0049] In one embodiment, the experimental animal is a rat, and the rat is 6 weeks to 20 weeks old.
[0050] It should be noted that since the physiology of rats is highly similar to that of humans, rats are the preferred model for the construction method of the present embodiment.
[0051] It should be noted that when the rat is too young, the eyeball is too small and has not fully developed, and the amount of material taken is too small, which is not conducive to detection; when the rat is too old, some cells in the eyeball appear to be aging, which is not conducive to the objective evaluation of changes in indicators, therefore, rats aged 6 weeks to 20 weeks are selected as experimental animal models.
[0052] It should be noted that rats aged 6 weeks to 20 weeks are selected, the size of the eyeball is suitable for experimental sampling, and the amount required for biochemical index detection is also met, which is more conducive to experimental operation.
[0053] In one embodiment, the experimental light source is ultraviolet light, blue light, or green light.
[0054] It should be noted that the shorter the wavelength, the higher the energy, so ultraviolet light, blue light, or green light with a short wavelength is selected as the experimental light source.
[0055] It should be noted that a single light source is used for visual fatigue experiments during the experiment, for example, ultraviolet light is selected alone for visual fatigue experiments.
[0056] In the present embodiment, the experimental animal grouping of the present embodiment is shown in Table 1 as follows.
[0057] Table 1 Experimental animal grouping
[0058]
[0059] In the embodiment, all normal groups are raised in an environment simulating daily light, with light intensity of 400 lux, and 10% concentration of physiological saline is dropped in the eyes of experimental animals before light in each experimental group every day.
[0060] After the light experiment is completed, the experimental animals are first detected in vivo.
[0061] In the embodiment, the intraocular pressure detection results of experimental animals in the embodiment are shown in Table 2 as follows.
[0062] Table 2 Intraocular pressure detection results
[0063]
[0064] Specifically, in the embodiment, the intraocular pressure of experimental animals is detected by Reichert tonometer.
[0065] In combination with Table 1 and Table 2, the following comparison results can be obtained:
[0066] Compared between MO1 and N1, under the conditions of same light period, humidity and dark damage period, the intraocular pressure increases with the increase of light intensity;
[0067] Compared between MO1 and MO2, under the conditions of same light intensity, humidity and dark damage period, the intraocular pressure increases with the increase of light period;
[0068] Compared between MO5 and N5, under the conditions of same light period, light intensity and dark damage period, the intraocular pressure increases with the decrease of humidity.
[0069] In summary, different light periods, different humidities and different light intensities all have effects on intraocular pressure, that is, have effects on visual fatigue.
[0070] In the embodiment, the visual detection results of experimental animals in the embodiment are shown in Table 3 as follows.
[0071] Table 3 Visual detection results
[0072]
[0073]
[0074] In combination with Table 1 and Table 3, the following comparison results can be obtained:
[0075] Compared between MO1 and N1, under the conditions of same light period, humidity and dark damage period, the visual acuity decreases with the increase of light intensity;
[0076] Compared with MO2, under the same light intensity, humidity and dark damage period, visual acuity decreases with the increase of light cycle;
[0077] Compared with MO5, under the same light cycle, light intensity and dark damage period, visual acuity decreases with the decrease of humidity.
[0078] It should be noted that the decrease of visual acuity is the key to the occurrence of visual fatigue. The visual acuity of a normal rat is generally 0.5cpd to 1.0cpd, and the visual acuity of animals with mild to severe visual fatigue decreases by about 0.1.
[0079] It should be noted that when the visual fatigue of the experimental animal is severely damaged to be irreversible, the visual acuity of the experimental animal cannot be detected, resulting in a visual acuity range detection value lower than 0.1. Compared with MO3, under the same light cycle, humidity and dark damage period, the visual acuity detection result of MO7 is lower than 0.1, indicating that the light intensity has caused irreversible visual fatigue damage to the experimental animal.
[0080] In summary, different light cycles, different humidities and different light intensities all have an impact on visual acuity, that is, an impact on visual fatigue.
[0081] In this embodiment, the TBUT test results of the experimental animals of this embodiment are shown in Table 4 as follows.
[0082] Table 4 TBUT test results
[0083]
[0084]
[0085] Specifically, 1% fluorescein sodium solution is dropped into the conjunctival sac of the animal, 1 drop, and waited for 1 min; observation and timing are carried out under the blue light of the slit lamp microscope.
[0086] It should be noted that fluorescein sodium staining can observe the damage of the corneal and conjunctival epithelium, the integrity of the tear film and detect the tear breakup time (TBUT).
[0087] Combining Table 1 and Table 4, the following comparison results can be obtained:
[0088] Compared with MO1, under the same light cycle, humidity and dark damage period, TBUT decreases with the increase of light intensity;
[0089] Compared with MO1, under the same light intensity, humidity and dark damage period, TBUT decreases with the increase of light cycle;
[0090] MO5 and N5 were compared, under the condition of the same light period, light intensity and dark damage period, TBUT decreased with the decrease of humidity.
[0091] In summary, different light periods, different humidities and different light intensities all have an impact on TBUT, that is, an impact on visual fatigue.
[0092] In this embodiment, the experimental animal Schirmer test results of this embodiment are shown in the following Table 5.
[0093] Table 5 Schirmer test results
[0094]
[0095]
[0096] Specifically, the tear detection phenol red cotton thread is selected for testing. The more tears, the longer the red color, and vice versa.
[0097] It should be noted that the higher the amount of tears represents the lower degree of eye dryness and the lower degree of visual fatigue.
[0098] Combining Table 1 and Table 5, the following comparison results can be obtained:
[0099] MO1 and N1 were compared, under the condition of the same light period, humidity and dark damage period, the length of the tear-soaked cotton thread decreased with the increase of light intensity;
[0100] MO1 and MO2 were compared, under the condition of the same light intensity, humidity and dark damage period, the length of the tear-soaked cotton thread decreased with the increase of light period;
[0101] MO5 and N5 were compared, under the condition of the same light period, light intensity and dark damage period, the length of the tear-soaked cotton thread decreased with the decrease of humidity.
[0102] In summary, different light periods, different humidities and different light intensities all have an impact on the length of the tear-soaked cotton thread, that is, an impact on visual fatigue.
[0103] In this embodiment, the experimental animal SOD test results of this embodiment are shown in the following Table 6.
[0104] Table 6 SOD test results
[0105]
[0106]
[0107] Specifically, superoxide dismutase (SOD) is selected for testing, and the degree of visual fatigue is reflected by the numerical value of the antioxidant index.
[0108] It should be noted that superoxide dismutase is an important member of the antioxidant enzyme system in the biological system, and superoxide dismutase can remove superoxide anion free radicals (O2 - ), and protect cells from damage. The main physiological cause of visual fatigue is that external stimulation causes increased oxidative stress in the eye, which breaks the internal balance and reduces antioxidant indicators.
[0109] It should be noted that the lower the numerical value of the antioxidant index, the higher the oxidative stress level of the experimental animal, and the more serious the degree of visual fatigue.
[0110] According to Tables 1 and 6, the following comparison results can be obtained:
[0111] Comparing MO1 and N1, under the same light cycle, humidity and dark damage period, SOD decreases with the increase of light intensity;
[0112] Comparing MO1 and MO2, under the same light intensity, humidity and dark damage period, SOD decreases with the increase of light cycle;
[0113] Comparing MO5 and N5, under the same light cycle, light intensity and dark damage period, SOD decreases with the decrease of humidity.
[0114] In summary, different light cycles, different humidities and different light intensities all affect SOD, that is, they affect visual fatigue.
[0115] In this embodiment, the GSH-Px test results of the experimental animals of this embodiment are shown in Table 7 as follows.
[0116]
[0117]
[0118] Specifically, glutathione peroxidase (GSH-PX) is selected for testing, and the degree of visual fatigue is reflected by the numerical value of the antioxidant index.
[0119] It should be noted that glutathione peroxidase is an important catalytic enzyme for decomposing hydrogen peroxide widely existing in the body. It specifically catalyzes the reduction reaction of hydrogen peroxide by reduced glutathione (GSH), and can play a role in protecting the structural and functional integrity of the cell membrane. The main physiological cause of visual fatigue is that external stimulation causes the oxidative stress in the eye to increase, so that the internal balance is broken and the antioxidant index decreases.
[0120] It should be noted that the lower the value of the antioxidant index, the higher the oxidative stress level of the experimental animal, and the more serious the degree of visual fatigue.
[0121] According to Tables 1 and 7, the following comparison results can be obtained:
[0122] Comparing MO1 and N1, under the same light period, humidity and dark damage period, GSH-Px decreases with the increase of light intensity;
[0123] Comparing MO1 and MO2, under the same light intensity, humidity and dark damage period, GSH-Px decreases with the increase of light period;
[0124] Comparing MO5 and N5, under the same light period, light intensity and dark damage period, GSH-Px decreases with the decrease of humidity.
[0125] In summary, different light periods, different humidities and different light intensities all have an impact on GSH-Px, that is, an impact on visual fatigue.
[0126] In this embodiment, the MDA test results of the experimental animals of this embodiment are shown in the following Table 8.
[0127]
[0128]
[0129] Specifically, MDA is a lipid peroxide, and the content of MDA in the experimental animals is detected in this embodiment.
[0130] It should be noted that the biological organism produces oxygen free radicals through enzyme systems and non-enzyme systems, and the oxygen free radicals can attack polyunsaturated fatty acids in biological membranes, induce lipid peroxidation, and thus generate lipid peroxides. Oxygen free radicals can cause cell damage through the decomposition products of lipid peroxides, so the amount of MDA tested can often reflect the degree of lipid peroxidation in the body, and indirectly reflect the degree of cell damage.
[0131] It should be noted that the degree of cell damage can also be reflected by testing other lipid peroxides, such as ketone, hydroxyl, carbonyl, hydroperoxyl or endoperoxide.
[0132] It should be noted that the MDA test is often combined with the SOD test, and the SOD index value indirectly reflects the body's ability to remove oxygen free radicals, while the MDA index value indirectly reflects the severity of the body's cells being attacked by free radicals. The results of the SOD test and the MDA test analysis are helpful to determine the development of visual fatigue.
[0133] Based on Tables 1 and 7, the following comparison results can be obtained:
[0134] Comparing MO1 and N1, under the same light period, humidity and dark damage period, MDA increases with the increase of light intensity;
[0135] Comparing MO1 and MO2, under the same light intensity, humidity and dark damage period, MDA increases with the increase of light period;
[0136] Comparing MO5 and N5, under the same light period, light intensity and dark damage period, MDA increases with the decrease of humidity.
[0137] In summary, different light periods, different humidities and different light intensities all have an impact on MDA, that is, they have an impact on visual fatigue.
[0138] Based on Tables 1 to 7, the following comparison results can be obtained:
[0139] Under the same light period, humidity and dark damage period, the increase of light intensity will increase the damage degree of visual fatigue;
[0140] Under the same light intensity, humidity and dark damage period, the increase of light period will increase the damage degree of visual fatigue;
[0141] Under the same light period, light intensity and dark damage period, the decrease of humidity will increase the damage degree of visual fatigue.
[0142] Although embodiments of the present application have been described, those skilled in the art can make various modifications and variations without departing from the spirit and scope of the present application, and such modifications and variations fall within the scope defined by the appended claims.
Claims
1. A method for constructing an animal model of visual fatigue, characterized by, The application relates to a method for preparing an experimental animal model of light-induced retinal damage. The experimental animal is placed in a dark environment for the first time for dark adaptation; The experimental animal is a rat; The experimental animal is randomly divided into a normal group and an experimental group, and the normal group and the experimental group are subjected to light exposure experiments; the normal group is bred in an environment simulating daily light exposure, and the experimental group is bred in an animal light damage device; the animal light damage device is placed in an environment with light intensity of 1000 lux to 5000 lux of an experimental light source; The experimental animal is subjected to an experiment for a light exposure cycle t, and the light exposure cycle t is 20 days to 40 days; The experimental group is placed in an experimental environment with humidity of 20%; During the light exposure cycle, 10% to 20% concentration of normal saline is dropped in the eyes of the experimental animal before light exposure every day; After the light exposure cycle, the experimental animal is first placed in a dark environment for the second time for dark adaptation; during the second time for dark adaptation, only maintenance feed is provided to the experimental animal to maintain life; The experimental animal is subjected to physical index detection.
2. The method of claim 1, wherein the method is performed for 1 to 7 days. The cycle of the second time for dark adaptation is 5 days to 15 days.
3. The method of constructing an animal model of visual fatigue according to any one of claims 1-2, wherein, The light exposure mode of the light exposure cycle is 12 hours / 12 hours light / dark cycle.
4. The method of constructing an animal model of visual fatigue according to any one of claims 1 to 2, wherein The age of the rat is 6 weeks to 20 weeks.
5. The method of claim 1-2, wherein, The experimental light source is ultraviolet light, blue light or green light.