CN115336553A - Construction method and application of open-angle glaucoma disease animal model - Google Patents

Abstract

A construction method and application of an open-angle glaucoma animal model simulate the occurrence of oxidative damage induced open-angle glaucoma from the pathogenesis of glaucoma, the animal modeling method has high success rate and has the following characteristics similar to human open-angle glaucoma: the angle of the house is open, the trabecular meshwork is oxidatively damaged, the aqueous humor outflow passage is fibrosis-changed, and the ocular hypertension is high. The invention can well solve the defects of large difference of modeling success rate and small amplitude of expansion existing in a hormone induction method, the modeling effect of the animal open-angle glaucoma model is superior to that of other existing open-angle animal models, and the animal open-angle glaucoma model has the advantages of simplicity, stability, short period and high repeatability.

Description

Construction method and application of an animal model of open-angle glaucoma

technical field

The invention relates to the technical field of disease animal models, in particular to a construction method and application of an open-angle glaucoma disease animal model.

Background technique

Glaucoma is the world's first irreversible blinding eye disease. It is estimated that by 2040, the number of patients will exceed 110 million. Among them, the onset of primary open-angle glaucoma is relatively hidden. If it is not diagnosed and treated in time in the early stage, it can lead to progressive loss of visual field and seriously affect the quality of life of the patient. Increased intraocular pressure leads to damage to retinal ganglion cells. At present, the pathogenesis of primary open-angle glaucoma is complex and not completely clear, but many studies have shown that oxidative damage is the main risk factor leading to it.

Nowadays, more open-angle glaucoma models are used, which are mainly divided into gene-induced and manipulation-intervention models according to the construction method. Gene-induced model animal models require high technical requirements, are expensive, and have a low success rate. At present, the more commonly used method is to operate the induced ocular hypertension model, which is induced by chemical reagents, such as dexamethasone and benzalkonium bromide. Using dexamethasone to model multi-simulating steroid glaucoma has a large difference in the success rate of modeling, while benzalkonium bromide applied to the eye is more suitable for the study of ocular surface drug toxicity. Currently, various animal models of rodent open-angle glaucoma have certain defects and cannot meet the current research needs of open-angle glaucoma. Therefore, it is necessary to seek a new modeling method.

Contents of the invention

In order to solve the defects and deficiencies of the existing open-angle glaucoma models, the present invention provides a construction method and application of an animal model of open-angle glaucoma disease, which is more similar to human open-angle glaucoma eye features, and the induction period is short, The method has a high success rate, is stable, and is easy to operate, and solves the problem that the hormone-induced method cannot fully simulate the ocular manifestations of human open-angle glaucoma.

The technical solution adopted in the present invention is: a method for constructing an animal model of open-angle glaucoma disease, constructed by any one of the following methods:

or

(2) 20 μL of 0.1% tert-butyl hydroperoxide (tBHP) was injected under the conjunctiva of the animal's ocular surface, the injection frequency was three times a week, and the interval between two injections was 1-2 days.

The construction period of the described animal model construction method is 1-28 days.

The animals are C57BL/6J mice or SD rats.

Application of tert-butyl hydroperoxide in the preparation of an inducing drug for constructing an animal model of open-angle glaucoma disease.

The inducing drug is eye drops or injections.

The concentration of tert-butyl hydroperoxide in the inducing drug is 0.1%.

The beneficial effects of the present invention are: the present invention provides a construction method and application of an animal model of open-angle glaucoma disease, which simulates the occurrence of open-angle glaucoma induced by oxidative damage from the pathogenic mechanism of glaucoma, and the success rate of the animal modeling method High, and has the following characteristics similar to human open-angle glaucoma: open chamber angle, oxidative damage to trabecular meshwork, fibrosis of aqueous humor outflow channel and high intraocular pressure. The present invention can well solve the drawbacks of the hormone induction method that the modeling success rate varies greatly and the increase is small. The modeling effect of the animal open-angle glaucoma model of the present invention is better than that of other open-angle glaucoma models currently available. Animal models have the advantages of simplicity, stability, short cycle time and high repeatability.

Description of drawings

Fig. 1 is a graph of intraocular pressure fluctuations in C57BL/6J mice administered with 0.1% tBHP for 28 days in Example 1.

Fig. 2 is a diagram of intraocular pressure fluctuations in SD rats in Example 2 using 0.1% tBHP instilled in the eyes for 28 days.

Figure 3 is a representative diagram of the anterior segment structure of C57BL/6J mice; Figure 3A is PBS eye drops of C57BL/6J mice; Figure 3B is 0.1% tBHP eye drops of C57BL/6J mice; Figure 3C is immunofluorescence showing C57BL The expression of TGF-β and α-SMA increased in /6J mice after eye drops of 0.1% tBHP compared with the control group.

Figure 4 is a representative diagram of the structure of the anterior segment of SD rats; Figure 4A is the eye drops of SD rats with PBS; Figure 4B is the eyes of SD rats with 0.1% tBHP; Figure 4C is the immunofluorescence display of SD rats with 0.1% tBHP Compared with the control group, the expressions of TGF-β and α-SMA increased after eye drops.

Detailed ways

The present invention will be further described below in conjunction with the drawings and the following embodiments. It should be understood that the drawings and the following embodiments are only used to illustrate the present invention rather than limit the present invention.

Example 1: Increased intraocular pressure in C57BL/6J mice after application of 0.1% tBHP eye drops

0.1% tBHP was administered to C57BL/6J mice. The results are shown in Figure 1. It was found that the intraocular pressure of the mice increased from day 2 after modeling and remained stable during the observation period of 4-28 days.

Embodiment 2: SD rat application of 0.1% tBHP after eye drop increases intraocular pressure

Eye drops of 0.1% tBHP were given to SD rats, and the results are shown in Figure 2. It was found that the intraocular pressure of the rats increased from the second day after modeling, and remained stable during the observation period of 4-28 days.

Example 3: The hardness of the trabecular meshwork increased after C57BL/6J mice were applied with 0.1% tBHP eye drops

0.1% tBHP was administered to C57BL/6J mice, and the results are shown in Figure 3. The chamber angle of the mice was open, and the trabecular meshwork structure was more compact than that of the control group, and the fibrosis-related proteins TGF-β, α-SMA expression increased.

Embodiment 4: SD rats apply 0.1% tBHP eyedrops to increase the hardness of trabecular meshwork

0.1% tBHP eye drops were given to SD rats, the results are shown in Figure 4, the chamber angles of the rats were open, and the structure of the trabecular meshwork was more compact than that of the control group, and the fibrosis-related proteins TGF-β, α- SMA expression increased.

in conclusion

The present invention simulates the occurrence of open-angle glaucoma induced by oxidative damage from the pathogenic mechanism of glaucoma, and the pathophysiological changes of the eyes are similar to those of human open-angle glaucoma: the chamber angle is opened, the trabecular meshwork is oxidatively damaged, and fibrosis changes are increased. Lead to increased hardness, increased intraocular pressure. By using tert-Butylhydroperoxide (tBHP) to induce oxidative damage to the trabecular meshwork in mice, fibrosis of the aqueous humor outflow channel and high intraocular pressure, the animal open-angle glaucoma model has a high success rate, The modeling cycle is short, the intraocular pressure increases significantly, and the advantages are simple and stable.

Notes to all technical personnel: Although the present invention has been described according to the above-mentioned specific embodiments, the inventive idea of the present invention is not limited to this invention, and any modification using the inventive idea will be included in the scope of protection of this patent.

The above descriptions are only preferred implementations of the present invention, and the scope of protection of the present invention is not limited to the above-mentioned embodiments, and all technical solutions under the idea of the present invention belong to the scope of protection of the present invention. It should be pointed out that for those skilled in the art, some improvements and modifications without departing from the principle of the present invention should also be regarded as the protection scope of the present invention.

Claims (6)

1. A construction method of an open-angle glaucoma disease animal model, characterized in that, it is constructed by any one of the following methods: or (2) 20 μL of 0.1% tert-butyl hydroperoxide (tBHP) was injected under the conjunctiva of the animal's ocular surface, the injection frequency was three times a week, and the interval between two injections was 1-2 days. 2. The construction method according to claim 1, characterized in that, the construction period of the animal model construction method is 1-28 days. 3. The construction method according to claim 1, wherein the animal is a C57BL/6J mouse or a SD rat. 4. The application of tert-butyl hydroperoxide in the preparation of an inducing drug for constructing an animal model of open-angle glaucoma disease. 5. The application according to claim 5, characterized in that the inducing drug is eye drops or injections. 6. The application according to claim 5, characterized in that the concentration of tert-butyl hydroperoxide in the inducing drug is 0.1%.

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