Method for establishing a closed-angle glaucoma animal model

Abstract

The application discloses a method for establishing an angle-closure glaucoma animal model. The method comprises the following steps: performing mydriasis on an eye of an animal model after general anesthesia, performing local anesthesia on an eye surface of the model eye after 10 minutes; after the pupil of the model eye is reduced, eye gel is added to the eye surface; a semiconductor laser is used to uniformly hit a plurality of laser points at an iris margin; and eye drops are added to the eye surface after laser treatment. The method uses a 532nm laser to hit the iris margin, causes local edema of the iris margin, narrows a passage through which aqueous humor flows from the posterior chamber into the anterior chamber, and increases the pressure of the posterior chamber. The method can keep the high intraocular pressure of the established angle-closure glaucoma animal model between 20-30mmHg, the success rate is 70%, the high intraocular pressure can be maintained for more than one week, no wound is caused on the eye of the animal model, there is no inflammatory reaction on the eye surface, the cornea has no edema and new blood vessels, and the operation of the model can be completed within 10 minutes.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of animal models, and particularly relates to a method for establishing an animal model of closed-angle glaucoma. BACKGROUND

[0002] Glaucoma is a group of diseases with optic nerve atrophy and visual field defects as common characteristics, and the main risk factor is the pathological intraocular pressure (IOP) increase. The compression and damage of intraocular tissues, especially the optic nerve head (ONH) of the posterior segment of the eye, caused by high intraocular pressure, lead to optic atrophy and visual field loss, which is the main cause of glaucoma blindness. At present, the main treatment for glaucoma is to reduce intraocular pressure by drugs or surgery to prevent or delay the progression of the disease. There is no report on the method for establishing an animal model of closed-angle glaucoma at home and abroad. Therefore, it is of great significance to establish an effective animal model of closed-angle glaucoma for the study of the pathogenesis of glaucoma and the search for new treatment methods and drugs to prevent, treat or delay the course of glaucoma.

[0003] The commonly used animal models of glaucoma at present mainly include: 1) posterior chamber injection of alpha-chymotrypsin or carbomer high intraocular pressure model: the disadvantages are that it can cause obvious inflammatory reaction, the intraocular pressure rises suddenly, the maintenance time is short, the success rate is low, and many complications such as lens turbidity, dislocation, anterior chamber exudation, corneal edema and lens atrophy are easy to cause. 2) anterior chamber injection of sodium hyaluronate high intraocular pressure model: the disadvantages are that it needs to be repeatedly injected to maintain high intraocular pressure, and the intraocular pressure does not rise obviously; usually, simple injection of sodium hyaluronate into the anterior chamber can only maintain high intraocular pressure for 5 days, and if long-term high intraocular pressure is needed, repeated injection of sodium hyaluronate every 5 days is required. 3) laser photocoagulation of sclera or laser photocoagulation of trabecular meshwork high intraocular pressure model: the operation is difficult, the high intraocular pressure is maintained for a short time and the intraocular pressure is unstable, repeated laser photocoagulation is needed to maintain high intraocular pressure for a long time, and this method also causes intraocular hyperemia and inflammatory reaction. 4) anterior chamber injection of microspheres to establish a chronic high intraocular pressure model: polystyrene microspheres are injected into the anterior chamber to block the outflow of anterior chamber fluid, causing high intraocular pressure. Due to the influence of gravity, the microspheres injected in the chamber angle are not evenly distributed, resulting in unstable intraocular pressure rise, and repeated injection of microspheres is needed to maintain high intraocular pressure for a long time.

[0004] At present, there is no method for establishing an animal model of closed-angle glaucoma. SUMMARY

[0005] In order to solve the above-mentioned technical problems, the purpose of the present application is to establish a method for establishing an animal model of closed-angle glaucoma caused by iris bulging induced by pupillary block mechanism.

[0006] The technical scheme adopted by the present application is: a method for establishing a closed-angle glaucoma animal model, comprising the following steps:

[0007] Step 1: Mydriasis is performed on the animal model eye, and after 10 minutes, local anesthesia is performed on the ocular surface of the model eye.

[0008] Step 2: After the pupil of the model eye is reduced, eye gel is added to the ocular surface.

[0009] Step 3: A semiconductor laser is used to uniformly hit a plurality of laser points at the iris margin.

[0010] Step 4: After laser treatment, eye drops are added to the ocular surface.

[0011] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 1, the mydriasis is: mydriasis is performed by adding pilocarpine eye drops to the animal model eye.

[0012] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 1, the local anesthesia is: local anesthesia is performed on the ocular surface of the model eye by adding obicaine hydrochloride eye drops.

[0013] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 2, the eye gel is sodium hyaluronate eye gel.

[0014] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 2, after the eye gel is added to the ocular surface, a transparent cover glass is used to flatten the cornea.

[0015] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 3, the semiconductor laser is a semiconductor laser with a laser wavelength of 532 nm.

[0016] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 3, uniformly hitting a plurality of laser points at the iris margin is: uniformly hitting 40-80 laser points at the iris margin; the laser diameter is 50-100 um, and the laser intensity is 100-150 mV.

[0017] Further, in the above method for establishing a closed-angle glaucoma animal model, in step 4, after laser treatment, levofloxacin eye drops are added to the ocular surface.

[0018] Further, in the above method for establishing a closed-angle glaucoma animal model, the animal is a rabbit or a mouse.

[0019] The present application provides the use of the establishment method in establishing a closed-angle glaucoma animal model.

[0020] The beneficial effects of the present application are:

[0021] 1、The method for establishing an animal model of closed-angle glaucoma provided by the present application can reduce the pupil of the model eye before laser ablation of the iris, which can make the iris expand, facilitate laser ablation operation and uniform distribution of laser sites, and more importantly, make the iris margin closer to the anterior lens capsule, increase the pupil blockage effect of the model eye, increase the intraocular pressure, form iris bulging, and effectively increase the pressure in the posterior chamber by comprehensively considering the above two points. The iris bulges towards the cornea to achieve the purpose of occluding the anterior chamber angle and increasing the intraocular pressure.

[0022] 2、The method for establishing an animal model of closed-angle glaucoma provided by the present application selects a 532 mn semiconductor laser because the laser is stable, the cost is low, the controllable range of laser energy is wide, the intensity of 100-150 mV laser is weak, the damage to other parts of the eyeball is small, and only a small range of edema of the iris margin is formed after laser ablation of the iris margin with a laser diameter of 50-100 um, without causing inflammation of other tissues in the eye.

[0023] 3、The method for establishing an animal model of closed-angle glaucoma provided by the present application can prevent damage to the corneal epithelial cell layer when the laser passes through the cornea by adding sodium hyaluronate eye gel to the eye before laser ablation of the iris margin, and can reduce the inflammatory reaction in the cornea after laser ablation of the iris margin; the use of a cover glass to flatten the cornea during laser ablation can reduce the scattering and refraction of laser rays caused by the curvature of the cornea when the laser passes through the cornea, reduce the weakness and dispersion of laser energy, and further reduce the damage to other tissues in the eyeball caused by the laser, so that the laser can be more concentrated and accurately ablated to the predetermined position of the iris margin.

[0024] 4、The method for establishing an animal model of closed-angle glaucoma provided by the present application can reduce the stress inflammatory reaction of the cornea to the laser by adding levofloxacin eye drops to the ocular surface after laser ablation of the iris margin, and ensure the normal physiological structure and function of the cornea.

[0025] 5、The method for establishing an animal model of closed-angle glaucoma provided by the present application can maintain the high intraocular pressure at 20-30 mmHg through a single operation after laser ablation of the iris margin. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 is a schematic diagram of laser ablation of the iris margin of closed-angle glaucoma;

[0027] Among them, A is the laser ablation area of the iris of the model eye, and the black concentric circle is the laser area; B is the distribution position and shape of the iris of the model eye before laser modeling; C is the distribution position and shape of the iris of the model eye after laser modeling.

[0028] Figure 2 Figure 1 shows the anterior segment examination pictures of the control eye and the model eye;

[0029] Figure 1 shows the anterior segment examination pictures of the control eye and the model eye;

[0030] Figure 3 Figure 2 shows the fundus examination pictures of the control eye and the model eye;

[0031] Figure 2 shows the fundus examination pictures of the control eye and the model eye;

[0032] Figure 4 Figure 3 shows the intraocular pressure change statistics of the control eye and the model eye within 7 days after modeling. DETAILED DESCRIPTION

[0033] Example 1

[0034] (I) The method for establishing an animal model of closed-angle glaucoma

[0035] For example, a New Zealand rabbit raised for 12 weeks is used to establish an animal model of closed-angle glaucoma, and the right eye is used as the model eye, and the left eye is not treated as the normal control eye. The method includes the following steps:

[0036] Step 1: Mydriasis is performed on the animal model eye, and 10 minutes later, local anesthesia is performed on the ocular surface of the model eye.

[0037] As a preferred embodiment, the mydriasis is that the animal model eye is dripped with pilocarpine eye drops for mydriasis.

[0038] As a preferred embodiment, the local anesthesia is that the ocular surface of the model eye is dripped with ophthalmic hydrochloric acid for local anesthesia.

[0039] As an exemplary illustration, in this embodiment, the New Zealand rabbit model eye (right eye) after general anesthesia is dripped with pilocarpine eye drops for mydriasis, and 10 minutes later, ophthalmic hydrochloric acid is dripped for local anesthesia on the ocular surface.

[0040] Step 2: After the pupil of the model eye is reduced, ophthalmic gel is dripped on the ocular surface.

[0041] As a preferred embodiment, the ophthalmic gel is sodium hyaluronate ophthalmic gel.

[0042] As a preferred embodiment, after the ophthalmic gel is dripped on the ocular surface, a transparent cover glass is used to flatten the cornea.

[0043] As an example, in this embodiment, after the pupil of the model eye shrinks, sodium hyaluronate ophthalmic gel is applied to the ocular surface, and then a transparent coverslip is used to flatten the cornea.

[0044] Step 3: Use a semiconductor laser to uniformly amplify several laser points at the edge of the iris.

[0045] Preferably, the semiconductor laser is a semiconductor laser with a wavelength of 532 nm.

[0046] As a preferred method, uniformly agitating several laser points at the edge of the iris involves a total of 40-80 laser points at the edge of the iris, with a laser diameter of 50-100 μm and a laser intensity of 100-150 mV.

[0047] As an example, in this embodiment, a semiconductor laser with a wavelength of 532nm is used to uniformly agitate 40-80 laser points at the edge of the iris; the laser diameter is 50-100um and the laser intensity is 100-150mV.

[0048] Step 4: Apply eye drops to the ocular surface after laser treatment.

[0049] Preferably, the eye drops are levofloxacin eye drops.

[0050] As an example, in this embodiment, levofloxacin eye drops are applied to the ocular surface after laser treatment.

[0051] (II) Testing

[0052] 1. For example Figure 2 As shown, slit-lamp ocular surface testing was performed on the model eyeballs 1, 3, and 7 days after modeling. The anterior chamber depth and anterior chamber angle width of the model were observed at each time point using a slit lamp. On the 3rd and 7th days, the anterior chamber depth of the model became shallower, and the iris bulged, resulting in a narrower anterior chamber angle.

[0053] 2. For example Figure 3 As shown, fundus examinations were performed on the model eyes 1, 3 and 7 days after modeling. Fundus photography revealed reduced blood supply to the model eyes on the 3rd and 7th days.

[0054] 3. For example Figure 4 As shown, intraocular pressure was measured in the model eye and the blank control eye after modeling. The measurements were taken for 7 consecutive days. It was found that the intraocular pressure of the model eye was 20-30 mmHg, which was 12±2.4 mmHg higher than that of the control eye.

[0055] From the above experiment, the method of the application uses 532nm laser to hit the iris margin, causes local edema of the iris margin, narrows the passage of aqueous humor from the posterior chamber into the anterior chamber, increases the pressure in the posterior chamber, pushes the iris to the cornea direction under the pressure of the aqueous humor in the posterior chamber, and further changes the anterior chamber angle, forms the iris bulging caused by the pupil block mechanism, can simulate the pathogenesis and symptoms of the clinical angle closure glaucoma patients. The method of the application can make the established angle closure glaucoma animal model keep the high intraocular pressure between 20-30mmHg, the success rate is 70%, the high intraocular pressure can be maintained for more than 1 week, no wound is caused to the animal model eye, no inflammatory reaction, no corneal edema and neovascularization, the operation of the model can be completed within 10 minutes. The construction method provided by the application can be applied to the pathogenesis research and preclinical research of treatment drugs of angle closure glaucoma.

[0056] Embodiment 2 application

[0057] (I) the establishment method of the chronic angle closure glaucoma animal model

[0058] Taking the New Zealand rabbits raised for 12 weeks as an example, the chronic angle closure glaucoma animal model is established. The method comprises the following steps:

[0059] Step 1: after the New Zealand rabbit model eye (right eye) is anesthetized, the model eye is dropped with pilocarpine eye drops for mydriasis, and after 10 minutes, the eye surface is locally anesthetized by adding hydrochloric acid obcaid eye drops.

[0060] Step 2: after the pupil of the model eye is reduced, the eye surface is dropped with sodium hyaluronate eye gel, and then the cornea is flattened with a transparent cover glass.

[0061] Step 3: a semiconductor laser with a wavelength of 532nm is used to hit 40 laser points uniformly at the iris margin; the laser diameter is 100um, and the laser intensity is 100mV.

[0062] Step 4: after the laser treatment, the eye surface is dropped with levofloxacin eye drops.

[0063] The intraocular pressure detection results are shown in Table 1.

[0064] (II) the establishment method of the acute angle closure glaucoma animal model

[0065] Taking the New Zealand rabbits raised for 12 weeks as an example, the acute angle closure glaucoma animal model is established. The method comprises the following steps:

[0066] Step 1: after the New Zealand rabbit model eye (right eye) is anesthetized, the model eye is dropped with pilocarpine eye drops for mydriasis, and after 10 minutes, the eye surface is locally anesthetized by adding hydrochloric acid obcaid eye drops.

[0067] Step 2: After the pupil of the model eye is reduced, sodium hyaluronate eye gel is added to the ocular surface, and then the cornea is flattened with a transparent cover glass.

[0068] Step 3: A semiconductor laser with a wavelength of 532 nm is used to uniformly strike 80 laser points at the iris margin; the laser diameter is 50 um, and the laser intensity is 150 mV.

[0069] Step 4: After laser treatment, levofloxacin eye drops are added to the ocular surface.

[0070] The intraocular pressure detection results are shown in Table 1.

[0071] Table 1

[0072]

Claims

1. A method for establishing an animal model of closed-angle glaucoma, characterized in that, The method comprises the following steps: Step 1: Mydriasis is performed on the animal model eye, and after 10 minutes, topical anesthesia is performed on the ocular surface of the model eye; Step 2: After the pupil of the model eye is reduced, the ocular surface is dripped with eye gel; Step 3: Semiconductor laser is used to uniformly hit a plurality of laser points at the iris margin; Step 4: After laser treatment, eye drops are dripped on the ocular surface.

2. The method for establishing a closed-angle glaucoma animal model according to claim 1, characterized in that, In step 1, the mydriasis is that pilocarpine eye drops are dripped into the animal model eye to perform mydriasis.

3. The method for establishing an angle-closure glaucoma animal model according to claim 1, characterized in that, In step 1, the topical anesthesia is that ophthalmic ointment is dripped on the ocular surface of the model eye to perform topical anesthesia.

4. The method for establishing an angle-closure glaucoma animal model according to claim 1, characterized in that, In step 2, the eye gel is sodium hyaluronate eye gel.

5. The method for establishing an angle-closure glaucoma animal model according to claim 1, characterized in that, In step 2, after the ocular surface is dripped with eye gel, the cornea is flattened with a transparent cover glass.

6. The method for establishing an angle-closure glaucoma animal model according to claim 1, characterized in that, In step 3, the semiconductor laser is a semiconductor laser with a laser wavelength of 532 nm.

7. The method for establishing an angle-closure glaucoma animal model according to claim 1, characterized in that, In step 3, uniformly hitting a plurality of laser points at the iris margin is that 40-80 laser points are uniformly hit at the iris margin; the laser diameter is 50-100 um, and the laser intensity is 100-150 mV.

8. The method for establishing an angle-closure glaucoma animal model according to claim 1, characterized in that, In step 4, after laser treatment, levofloxacin eye drops are dripped on the ocular surface.

9. The method for establishing a closed-angle glaucoma animal model according to any one of claims 1-8, wherein, The animal is a rabbit or a mouse.

10. Use of the establishment method in any one of claims 1-8 in establishing an animal model of closed-angle glaucoma.

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