WO2017181835A1 - Method for establishing chronic ocular hypertension animal model - Google Patents

Abstract

A method for establishing a chronic ocular hypertension animal model. The method comprises: creating a glaucoma animal model having an elevated ocular pressure and damaged optic nerves by surgically inserting a material to block off an outflow pathway for an aqueous humor, such that the elevated ocular pressure is maintained and stabilized, and can be obtained and manipulated more easily. The obtained model has advantages such as a stable elevation and small fluctuations of the ocular pressure, the high ocular pressure can be maintained for a long time and the target ocular pressure can be controlled. A chronic ocular hypertension animal model can be made by using the method, and an ocular pressure thereof can be adjusted and controlled according to different degrees of blockage of the Schlemm's canal; the blockage of the Schlemm's canal can be completed by selecting fiber ducts of different specifications and sizes on the basis of different diameters of the Schlemm's canals in different types of animals, thereby generating chronic ocular hypertension animal models for multiple types of animals. The method enables the establishment of clinically compatible model-making animals and model-making methods for research into the mechanism of chronic ocular hypertension glaucoma, and provides a foundation for the mechanism of optic nerve damage in chronic glaucoma.

Description

 Method for establishing chronic high intraocular pressure animal model

 [0001] The present invention relates to the field of medical animal models, and in particular to a method for establishing a chronic high intraocular pressure animal model.

 [0002] Glaucoma is an irreversible optic nerve degenerative disease, the second blind eye disease in the world. The complex pathogenesis is still not clear. Although high intraocular pressure is not the only factor for glaucomatous visual impairment, high intraocular pressure is one of the most dangerous factors in the development and development of glaucomatous optic nerve damage. In the field of glaucoma research, animal models have become an important research tool. The glaucoma animal model plays an important role in better understanding the mechanism of injury and treatment of glaucomatous optic neuropathy. High intraocular pressure is the most important risk factor for glaucoma optic nerve damage. Therefore, most animal models of glaucoma research still induce high intraocular pressure in animals, and then study the physiological, pathological and pharmacological aspects of glaucoma. The ideal experimental animal model requires: The construction method is simple, the cost is low, the feeding and control are easy, the elevated intraocular pressure is stable, and the pathological changes characteristic of the retinal optic nerve occur.

 [0003] The animals currently used to make glaucoma models are lower mammals, mainly rabbits, mice, and monkeys. Although the mouse is easy to be transformed and propagated by transgenic technology, and the structure of the mouse eye is similar to the human eye in many aspects, such as trabeculae, Schlemm tube, ciliary body, retinal blood vessels, etc., the mouse eye is relatively small, which is not conducive to surgical operation. . The laser-induced high intraocular pressure model of monkeys is currently recognized as the most ideal animal model of high intraocular pressure. However, due to its high cost and complicated operation, it is not widely used in China. Although there is a morphological difference between the rabbit eye and the angle structure of the human eye, the aqueous outflow channel is the same, and there is continuous endothelial cell lining. Rabbits are the most commonly used experimental animals for making glaucoma models because they are inexpensive, docile, easy to obtain and operate.

[0004] According to the mechanism of high intraocular pressure formation and drug action sites, it is divided into anterior chamber angle (and trabecular) type, such as anterior chamber injection of hyaluronic acid, methyl cellulose, blood cells, compound carbomer solution, posterior chamber Injection, chymotrypsin, laser photocoagulation trabecular meshwork, topical cortisol and other methods; effect on the scleral aqueous reflux vein type such as the common scleral surface intravenous hypertonic saline method (Morre-Morrison model), burning sclera surface The aqueous humor drainage method (Shareef-Sharma model) and improved methods; in addition, there are some special species Hairy mice and transgenic techniques construct glaucoma models, such as DBA/2J and AKXD-28/Ty pure mice, natural glaucoma models constructed using gene targeting techniques. Peer to evaluate the pathological changes of retinal ganglion cells ( _RGCs ), optic nerve and retinal function in various models, evaluate the advantages and disadvantages of various models, and better construct in the study of chronic high intraocular pressure glaucoma mechanism. The clinical model and animal modeling methods provide a basis for further study of the mechanism of chronic high intraocular pressure glaucoma and provide new thinking for its treatment.

 technical problem

 [0005] Although there are many methods for making high intraocular pressure animal models, most of the current glaucoma animal models are acute high intraocular pressure models, although the intraocular pressure is increased, but the high intraocular pressure is kept short, and the fluctuation is large. It is difficult to simulate the characteristic optic nerve damage of glaucoma.

 Problem solution

 Technical solution

 [0006] In order to overcome the defects of the prior art, the present invention provides a method for establishing a chronic high intraocular pressure animal model.

[0007] The technical solution adopted by the present invention is: A method for establishing a chronic high intraocular pressure animal model, comprising the following steps: (1) binding the model animal to the operating table, disinfecting the eye towel, placing the tissue Routine, flush the conjunctival sac, and perform local anesthesia under the conjunctiva of the operation eye, and fix the suture of the upper rectus muscle; (2) According to the direction of the dial of the clock, cut the coronal conjunctiva from 11 o'clock to 1 o'clock, to 12 o'clock A shallow scleral flap with a size of 4*4 mm and a 1/3 scleral thickness with a limbus as the base, and a deep sclera of 3*3 mm in thickness and 2/3 scleral thickness below the superficial scleral flap. Flap, and peeling toward the cornea, find the Schlemm tube and cut the outer wall of the Schlemm tube; (3) Inject polymer hyaluronic acid into both ends of the Schlemm tube, pass the fiber catheter tube through the fistula, and azimuth at 9 or 3 o'clock. 1.0 mm at the edge, the anterior chamber puncture in the direction of the flat iris, gently press the posterior edge of the puncture, release the water to reduce the intraocular pressure, insert the fiber catheter into the end of the Schlemm tube after the anterior decompression puncture, and follow the Schlemm tube. 360 degrees or 2 70 degrees or 180 degrees or 90 degrees, cut the catheter, place it in the Schlemm to block the Schlemm tube, prevent the outflow of aqueous humor, suture the scleral flap, inject the balanced salt solution into the anterior chamber puncture, suture the conjunctival flap, and give anti-inflammatory drugs, finally Establish an animal model of elevated intraocular pressure.

[0008] In the step (1), the conjunctival sac is washed with a diluted iodophor solution. [0009] In the step (1), 0.4 ml of 2% lidocaine was used for subconjunctival local anesthesia.

[0010] In the step (1), a polymer hyaluronic acid is injected into both ends of the Schlemm tube by using a 30G needle.

[0011] The fiber conduit in the step (3) comprises a cylindrical solid conduit, and the conduit has a spherical structure at one end.

 [0012] The fiber conduit is 10 cm long and has a diameter of 200 μm, and the spherical diameter of one end of the conduit is 250 μm.

[0013] The fiber conduit is made of a polypropylene material.

 Advantageous effects of the invention

 Beneficial effect

 [0014] The beneficial effects of the present invention are: The present invention provides a method for establishing a chronic high intraocular pressure animal model, which is a chronic glaucoma animal model in which the intraocular pressure is blocked by surgically placing a material, thereby causing an increase in intraocular pressure. The chronic ocular hypertension animal model produced by this method can be adjusted according to the range of occlusion Schl emm. It is also possible to select fiber guides of different sizes according to the Schlemm pipe diameter of different animal types, and complete the Schlemm tube blockage, thereby producing a chronic high intraocular pressure model of various animal types. In the study of chronic high intraocular pressure glaucoma mechanism, it is better to construct a model animal and modeling method that is compatible with the clinical, and provide a basis for the mechanism of optic nerve injury in chronic glaucoma.

 Brief description of the drawing

 DRAWINGS

 1 is a schematic view showing the structure of a fiber conduit of the present invention.

Embodiments of the invention

 [0016] The present invention will be further described with reference to FIG. 1. Operation steps: The operation requires a long 10 cm diameter 200 μηι solid fiber catheter made of polypropylene material, and the tip end of the catheter has a diameter of 250 μηι, which is partially expanded to prevent intubation. Damage to the Schlemm wall tissue.

[0017] The model animal limbs are tied to the operating table, the eye is routinely disinfected and spread, placed in a device, diluted with iodophor solution to wash the conjunctival sac, and 0.4 ml of 2% lidocaine is applied to the subconjunctival local anesthesia. After that, the upper rectus traction suture is fixed, and according to the direction of the dial of the clock, the conjunctiva is cut at the edge of 11 o'clock to 1 o'clock, and the size of the corneal margin is about 4*4 mm with the center of 12 o'clock as the center. 1/3 scleral thickness of the superficial scleral flap, A deep scleral flap with a size of about 3*3 mm and a thickness of about 2/3 scleral thickness is made under the superficial scleral flap. Peel off the cornea, find the Schlemm tube and cut the outer wall of the Schlemm tube, and inject the polymer hyaluronic acid (Healon GV) into the ends of the Schlem m tube with a 30G needle to make the catheter easily pass through the mouth. At the 1.0 mm point in the limbus at 9 or 3 o'clock, the anterior chamber puncture was performed in the direction of the flat iris. The posterior margin of the puncture was lightly pressed, and the right amount of aqueous humor was released to reduce the intraocular pressure. After the anterior chamber decompression puncture, the microcatheter was inserted into the Schlemm tube. Broken end, according to the required target intraocular pressure, respectively, follow the Schlemm tube 360 degrees or 270 degrees or 180 degrees or 90 degrees, cut the catheter, place it in Schlemm to block the Schlemm tube, prevent the outflow of water, resulting in A model of elevated intraocular pressure. Finally, 4-6 needles of the scleral flap were tightly sutured with a 10-0 polypropylene thread. The anterior chamber puncture was infused with a balanced salt solution, and the water-tight suture was observed as a non-leakage, and the conjunctival flap was sutured with a 10-0 polypropylene thread. Anti-inflammatory drugs were given after surgery to monitor intraocular pressure and anterior segment response.

 [0018] The present invention is a glaucoma animal model in which the intraocular water outflow channel is blocked by a surgically placed material, causing an increase in intraocular pressure to damage the optic nerve. The chronic high intraocular pressure animal model produced by this method can be based on the occlusion of Schlemm. The range is different for regulation. It is also possible to select fiber guides of different sizes according to the Schlemm pipe diameter of different animal types, and complete the Schlemm tube blockage, thereby producing a slow high intraocular pressure model of various animal types. . The animal models of rabbit eyes and monkey eyes use a fiber guide that can be used with a diameter of 10 cm and a diameter of 200 μm, and the tip of the catheter has a spherical diameter of 250 μm. Other animals can choose a catheter size that matches the Schlemm diameter. In the study of chronic high intraocular pressure glaucoma mechanism, it is better to construct a model animal and modeling method that is compatible with the clinical, and provide a basis for the mechanism of optic nerve injury in chronic glaucoma.

 The above description is only a preferred embodiment of the present invention, and the scope of protection of the present invention is not limited to the above embodiments, and all the technical solutions under the inventive concept belong to the protection scope of the present invention. It should be noted that those skilled in the art will be able to devise a number of improvements and modifications without departing from the principles of the invention.

Claims

Claim A method for establishing a chronic ocular hypertension animal model, comprising the steps of: (1) Tied the model animal to the supine position, disinfect the eyelids, place the device, and rinse the conjunctival sac After the local anesthesia under the conjunctiva of the operation eye, the upper rectus muscle traction suture is fixed;  (2) According to the direction of the dial of the clock, the cornea is cut at the edge of the 11 o'clock to the 1 o'clock edge, and the shallow layer of the 1/3 scleral thickness of the size 4*4 mm with the limbus as the base is centered at 12 o'clock. Scleral flap, under the superficial scleral flap, a deep scleral flap with a size of 3*3mm and a thickness of 2/3 scleral, and peeled off the cornea, find the Schlemm tube and cut the outer wall of the Schlemm tube;  (3) Inject polymer hyaluronic acid into both ends of the Schlemm tube, pass the fiber catheter catheter through the fistula, 1.0 mm in the corneal margin at 9 or 3 o'clock, and puncture the anterior chamber in the direction of the flat iris. At the trailing edge, release water to reduce intraocular pressure. After anterior chamber decompression puncture, insert the fiber catheter into the end of the Schlemm tube, and follow the Schlemm tube for 360 or 270 degrees or 180 degrees or 90 degrees. Cut the catheter and place it. Blocking the Schlemm tube in Schlemm, blocking the outside water, 3⁄4? , suture the scleral flap, inject the balanced salt solution into the anterior chamber puncture, suture the conjunctival flap, give anti-inflammatory drugs, and finally establish an animal model with elevated intraocular pressure. A method of establishing a chronic ocular hypertension animal model according to claim 1, wherein said step (1) is to wash the conjunctival sac with a diluted iodophor solution. The method for establishing a chronic ocular hypertension animal model according to claim 1, wherein said step (1) is performed by subcutaneous subconjunctival anesthesia using 0.4 ml of 2% lidocaine. A method of establishing a chronic ocular hypertension animal model according to claim 1, wherein said step (1) uses a 30G needle to inject high molecular hyaluronic acid into both ends of the Schlemm tube. A method of establishing a chronic ocular hypertension animal model according to claim 1, wherein said step (3) of the fiber conduit comprises a cylindrical solid catheter, and said catheter has a spherical structure at one end. A method for establishing a chronic ocular hypertension animal model according to claim 5, characterized in that The fiber catheter has a length of 10 cm and a diameter of 200 μm, and the spherical diameter of one end of the catheter is 250 μm.  [Claim 7] A method of establishing a chronic ocular hypertension animal model according to claim 5, wherein the fiber catheter is made of a polypropylene material.