Construction method of cataract animal model and light treatment experiment method

Abstract

The invention relates to the field of ophthalmology treatment, and discloses a cataract animal model construction method, which comprises the following steps: obtaining a plurality of 21-24-day-old SD rats; d-(+)-galactose drinking water with the concentration of 8% is prepared, the volume of the D-(+)-galactose drinking water is configured according to the weight of the SD rats, and 25 mL of the D-(+)-galactose drinking water is prepared every day for the SD rats with the configuration standard being 100 g; each SD rat is fed with D-(+)-galactose drinking water for 14 days, each SD rat is induced to form a cataract animal model, and then each SD rat is fed with pure water. According to the method, D-(+)-galactose with the concentration of 8% is selected to induce rats to form the cataract animal model, the formed cataract animal model is stable in health condition, the overall process of cataract is slow, and the method is suitable for observation before and after a light treatment experiment.

Description

Technical Field

[0001] The invention relates to the field of ophthalmic treatment, and in particular to a method for constructing a cataract animal model and a light treatment experimental method. Background Art

[0002] Cataract is a disease in which the lens of the eye becomes cloudy due to various reasons, causing visual impairment. There is no proven drug for the treatment of cataracts, and surgery is currently the main treatment, including phacoemulsification and femtosecond laser-assisted phacoemulsification. Although surgical treatment of cataracts can quickly improve the patient's vision, there is a risk of infection and bleeding, and the cost of surgery is relatively high.

[0003] In order to solve the problems of surgical treatment of cataracts, researchers have proposed a method of treating cataracts by emitting light of a specific wavelength to the affected eye through an LED light source (hereinafter referred to as phototherapy). Compared with surgical treatment, phototherapy has the advantages of low treatment cost, being completely non-invasive and painless, and easy to implement.

[0004] However, the effectiveness and safety of phototherapy have yet to be further studied. Therefore, it is necessary to create animal models suitable for phototherapy experiments to promote in-depth research on phototherapy. Summary of the invention

[0005] The invention provides a method for constructing a cataract animal model, so as to prepare an animal model suitable for phototherapy experiments.

[0006] In order to prepare an animal model suitable for phototherapy experiments, the present application provides a method for constructing a cataract animal model, comprising the following steps: an animal configuration step, obtaining 21-24 day old SD rats, the SD rats are of SPF grade, and weigh 55-65g; a drinking water configuration step, configuring 8% D-(+)-galactose drinking water, the volume of which is configured according to the weight of the SD rats, and the configuration standard is 25mL per day for a 100g SD rat; a galactose feeding step, feeding the SD rats with 8% D-(+)-galactose drinking water for 11-14 days to induce the SD rats to form a cataract animal model, wherein the feeding method is that the SD rats drink on their own; a pure water feeding step, after feeding the SD rats with 8% D-(+)-galactose drinking water for 11-14 days, switching to pure water to feed the SD rats.

[0007] In one embodiment, SD rats were fed with 8% D-(+)-galactose in drinking water for 14 days.

[0008] The present application also provides a light therapy experimental method, which comprises the following steps: using compound tropicamide eye drops to dilate the eye to be treated of the cataract animal model obtained by the above-mentioned construction method; inducing anesthesia of the cataract animal model with 3% isoflurane, and maintaining it in an anesthetized state with 1.5-2% isoflurane, and after the cataract animal model is anesthetized, placing it on a feedback heating pad to keep it warm, and the temperature of the heating pad is maintained between 35-36 degrees Celsius; using a therapeutic light source to irradiate the lens area of ​​the eye to be treated, and the irradiation time is 20 minutes; during the irradiation process, dripping artificial tears into the eye to be treated every 2 minutes.

[0009] This application has the following beneficial effects:

[0010] (1) By selecting D-(+)-galactose with a concentration of 8% to induce cataract animal model in rats, the formed cataract animal model has stable health status and the overall progression of cataract is relatively slow, which is suitable for before and after observation of light therapy experiments.

[0011] (2) The cataract animal model was induced by 8% D-(+)-galactose. The severity of cataract varies to a certain extent, which is conducive to verifying the effect of light therapy for cataracts of different degrees. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The above and other features of the present application will be more fully understood through the following description and the attached claims in conjunction with the accompanying drawings. It is understood that these drawings only depict several embodiments of the present application and should not be considered as limiting the scope of the present application. By using the accompanying drawings, the present application will be more clearly and detailed.

[0013] Figure 1 The quadrant division diagram of the rat eyeball of the present invention is shown.

[0014] Figure 2 Shown are comparison images of rat cataracts before and after treatment under slit lamp observation. DETAILED DESCRIPTION

[0015] In the following detailed description, reference is made to the accompanying drawings which form a part thereof. In the drawings, similar symbols generally represent similar components unless the context indicates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not intended to be limiting. Other embodiments may be employed and other changes may be made without departing from the spirit or scope of the subject matter of the present application. It is understood that various configurations, substitutions, combinations, and designs of various aspects of the present application generally described in this application and illustrated in the drawings may be made, all of which explicitly constitute a part of the present application.

[0016] The present application provides a method for constructing a cataract animal model, which comprises the following steps:

[0017] (1) Animal configuration: 21-24 day old SD rats (hereinafter referred to as rats) were obtained, wherein the rats were of SPF grade and weighed 55-65 g.

[0018] Specifically, 9 male rats, 8 female rats, and 17 rats were reared in the following environment: the room temperature of the animal room was 20°C to 26°C, the humidity was 40% to 70%, the minimum ventilation frequency was not less than 15 times / hour, the relative static pressure difference was not less than 10Pa, and the lighting was 12h / 12h day and night light and dark alternation. Rats were reared in a polyphenylene sulfone resin (PPSU) plastic box with bedding on the IVC cage. The specifications of the rearing box were: length × width × height = 437mm × 287mm × 230mm (with a drinking bottle and cage cover). The floor of the animal room was disinfected once a day, and wiped and disinfected with 0.2% 84 disinfectant or 2% sporicide every week. The two disinfectants were used alternately every week, and the cages, walls and ceilings were disinfected at least once a month. Rats were allowed to eat and drink freely. The animal cages were changed at least once a week, along with the bedding and drinking water. When drinking water, urine, loose stools, and other serious contamination of the bedding occurred, the feeding boxes were changed at any time. All cages were cleaned and sterilized with high-pressure steam before use. The feed used was standard SPF-grade rat maintenance feed.

[0019] (2) Drinking water preparation: Prepare 8% D-(+)-galactose drinking water. The volume of D-(+)-galactose drinking water is prepared according to the weight of the rat. The preparation standard is that for a rat of 100 g, 25 mL of D-(+)-galactose drinking water is prepared every day.

[0020] (3) Galactose feeding: Rats were fed with D-(+)-galactose drinking water for 14 days to induce cataract animal model in rats, wherein the feeding method was that the rats drank by themselves. In this embodiment, feeding with D-(+)-galactose drinking water for 14 days was taken as an example. In other embodiments, feeding with D-(+)-galactose drinking water for 11 days, 12 days or 13 days may be selected.

[0021] (4) Pure water feeding: After feeding the rats with D-(+)-galactose drinking water for 14 days, the feeding of D-(+)-galactose drinking water was stopped and the rats were fed with pure water instead.

[0022] The cataract animal model obtained by the above-mentioned model construction method was scored using the following cataract scoring criteria, and the scoring results are shown in Table 1.

[0023] Cataract scoring criteria: (1) The rat eyeball is divided into 12 quadrants. Figure 1As shown; (2) The score weight of each quadrant: the outermost circle (1, 2, 3, 4) has a maximum score of 4 points, the middle circle (5, 6, 7, 8) has a maximum score of 3 points, and the innermost circle (9, 10, 11, 12) has a maximum score of 2 points, with a total score of 36 points; (3) Scoring is based on the number and area of ​​vesicles in each quadrant and the proportion of cortical opacity in the quadrant. The more vesicles, the larger the area, and the greater the proportion of cortical opacity in the quadrant, the higher the score, and vice versa.

[0024] It should be noted that, in this example, a total of 17 rats participated in the model construction experiment. Table 1 lists the scores of 10 cataract animal models given by three ophthalmologists; the scores recorded in Table 1 are the total scores of the 12 quadrants in which each eyeball is divided.

[0025] Table 1 Scoring record of left and right eyes of rats

[0026]

[0027] Note: In the rat number, L represents the left eye of the rat, and R represents the right eye of the rat.

[0028] It can be seen from the scores in Table 1 that the cataract animal model formed in rats induced by D-(+)-galactose at a concentration of 8% has a relatively slow overall progression of cataracts without severe cataracts, and the severity of cataracts in various cataract animal models is different and there are certain differences.

[0029] The following is a description of the light therapy experiment conducted after the cataract animal model is constructed.

[0030] The light therapy experiment on the animal model of cataract includes the following steps:

[0031] (1) Tropicamide compound eye drops were used to dilate the eye to be treated in the cataract animal model.

[0032] (2) The cataract animal model was anesthetized by inducing anesthesia with 3% isoflurane and maintained in an anesthetized state with 1.5-2% isoflurane. After the cataract animal model was anesthetized, it was placed on a feedback heating pad to keep warm. The temperature of the heating pad was maintained between 35 and 36 degrees Celsius.

[0033] (3) A therapeutic light source is used to irradiate the lens area of ​​the eye to be treated for 20 minutes. During the irradiation process, artificial tears are dripped into the eye to be treated every 2 minutes to keep it moist.

[0034] We selected animal models of cataracts with different severity before treatment for example analysis. The experimental results are as follows: Figure 2 And as shown in Table 2.

[0035] Table 2 Record of pathological changes of cataract in animal model before and after treatment

[0036]

[0037] Note: The scores in Table 2 are the average scores of three ophthalmologists.

[0038] The present application uses D-(+) galactose with a concentration of 8% to induce the formation of a cataract animal model in rats. The formed cataract animal model has a stable health condition and a relatively slow overall progression of cataracts, which is suitable for before and after observations of phototherapy experiments. In addition, the cataract animal models induced have different degrees of cataract severity, which is beneficial for verifying the effect of phototherapy for cataracts of different degrees.

[0039] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for constructing an animal model of cataract, characterized in that: The steps include: Animal preparation step: obtaining 21-24 day old SD rats, the SD rats are of SPF grade and weigh 55-65 g; The step of preparing drinking water is to prepare 8% D-(+)-galactose drinking water, wherein the volume of the D-(+)-galactose drinking water is prepared according to the body weight of the SD rats, and the preparation standard is that for a 100 g SD rat, 25 mL is prepared per day; a galactose feeding step, feeding the SD rats with 8% D-(+)-galactose drinking water for 11-14 days to induce the SD rats to form the cataract animal model, wherein the feeding method is that the SD rats drink on their own; In the pure water feeding step, after the SD rats were fed with the D-(+)-galactose drinking water having a concentration of 8% for 11-14 days, the SD rats were fed with pure water instead.

2. The construction method according to claim 1, characterized in that: The SD rats were fed with 8% D-(+)-galactose in drinking water for 14 days.

3. A light therapy experimental method, characterized in that: The steps include: Using compound tropicamide eye drops to dilate the eye to be treated of the cataract animal model obtained according to the construction method of claim 1 or 2; Inducing anesthesia of the cataract animal model with 3% isoflurane and maintaining it in an anesthetized state with 1.5-2% isoflurane. After the cataract animal model is anesthetized, it is placed on a feedback heating pad for warmth preservation, and the temperature of the heating pad is maintained between 35-36 degrees Celsius; The lens area of ​​the eye to be treated is irradiated with a therapeutic light source for 20 minutes; during the irradiation process, artificial tears are dripped into the eye to be treated every 2 minutes.

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