CN-121971196-A - Construction method of retinal vein occlusion animal model

Abstract

The invention provides a construction method of a retinal vein occlusion animal model, and belongs to the technical field of medicines. Firstly, anaesthetizing the surface of an eyeball of an anaesthetized non-human primate, establishing two scleral tunnels for the puncture of the eyeball after disinfection, respectively inserting an illumination optical fiber and an intraocular laser optical fiber into the two scleral tunnels, then injecting a photosensitizer through a limb vein, and immediately performing laser photocoagulation on retinal veins under the illumination of the illumination optical fiber after injection to obtain the retinal vein occlusion animal model. The invention can successfully construct a retinal vein occlusion model, and secondary macular edema, the time required for vascular recanalization is long, the retinal hemorrhage after retinal vein occlusion can be maintained for 4-5 weeks, the sign of the retinal hemorrhage is close to that of human beings, the retinal vein occlusion model has clinical drug responsiveness, the clinical research value of the retinal vein occlusion model is shown, and a good research model is provided for the research on the aspects of etiology, pathology and therapeutics.

Inventors

• HU ZHENGTAO
• FENG YULIANG
• YANG JIAYI
• YANG KE
• WANG XUEWEI
• Dou shuai

Assignees

• 成都华西海圻医药科技有限公司

Dates

Publication Date

20260505

Application Date

20251229

Claims (10)

1. 1. A construction method of a retinal vein occlusion animal model is characterized by comprising the following steps of firstly conducting surface anesthesia on an eye of an anesthetized non-human primate, then conducting disinfection on eye skin, eyelid margin and conjunctival sac, then establishing two scleral tunnels for eyeball puncture, respectively inserting an illumination optical fiber and an intraocular laser optical fiber into the two scleral tunnels, then injecting a photosensitizer through limb vein, and immediately conducting laser photocoagulation on retinal vein under illumination of the illumination optical fiber after injection, thus obtaining the retinal vein occlusion animal model.
2. 2. The construction method according to claim 1, wherein the injection of the photosensitizer is a single injection, and the dosage is 15-25mg/kg.
3. 3. The method of claim 2, wherein the photosensitizer is used in an amount of 20mg/kg.
4. 4. A method of construction according to any one of claims 1 to 3, wherein the photosensitizer comprises mangash, erythrosine B or verteporfin.
5. 5. The method of claim 1, wherein the non-human primate comprises a cynomolgus monkey or a rhesus monkey.
6. 6. The method of claim 1, wherein the exposure time of the laser is 0.1-0.2s, the laser power is 100-200mW, and the laser wavelength is 530-535nm.
7. 7. The method of claim 6, wherein the exposure time of the laser is 0.15s, the laser power is 160mW, and the laser wavelength is 532nm.
8. 8. The method according to claim 1, wherein the laser photocoagulation is performed by laser photocoagulation of primary and/or secondary branches of the retinal vein and/or laser photocoagulation of primary and/or secondary branches of the subretinal vein.
9. 9. The construction method according to claim 8, wherein the laser photocoagulation comprises performing laser photocoagulation on primary and secondary branches of an upper retinal vein and primary and secondary branches of a lower retinal vein, performing photocoagulation from a proximal end to a distal end from 0.5-1.5 optic disc diameters, wherein the primary branch vein receiving photocoagulation has a length of 3.5-4.5 optic disc diameters, and the secondary branch vein has a length of 1.0-2.0 optic disc diameters, avoiding accompanying arteries and macula fovea, and wherein the secondary branch vein is a secondary branch vein facing the macula area.
10. 10. The method according to claim 9, wherein the laser photocoagulation is performed from the proximal end to the distal end from 1 disc diameter to the optic disc, the length of the primary branch vein receiving the photocoagulation is 4 disc diameters, the length of the secondary branch vein is 1.5 disc diameters, the concomitant artery and the macular fovea are avoided, and the secondary branch vein is the secondary branch vein facing the macular area.

Description

Construction method of retinal vein occlusion animal model Technical Field The invention belongs to the technical field of medicines, and particularly relates to a construction method of a retinal vein occlusion animal model. Background Retinal Vein Occlusion (RVO) is the second most vascular disease that leads to vision loss. RVOs can be classified into central venous occlusion (CRVO) and Branch RETINAL VEIN Occlusion (BRVO) according to retinal vein occlusion sites. Clinically, retinal vein occlusion often induces complications such as macular edema and neovascularization, leading to vision impairment and blindness. The prior RVO animal model is mostly constructed by adopting rodents (such as rats, mice) or rabbits and other animals through methods of laser direct photocoagulation, vessel ligation, drug induction and the like. However, these models have obvious limitations such as small eyeballs, no macula structure, difficulty in simulating macular edema in humans, large differences between rabbit retinal vascular systems and humans, and easiness in causing retinal thermal injury, high vascular re-circulation rate or inconsistent formation mechanisms of human RVO. Non-human primates (e.g., cynomolgus, rhesus) are considered ideal model animals because of their high similarity to humans in retinal anatomy, blood supply system, and macular characteristics. The existing RVO model building method mainly comprises a vascular ligation method, a diathermy method, an endothelin-1 (ET-1) injection method in a vitreous cavity, a laser photocoagulation closed retina vein method and a photosensitive medicine intravenous injection combined laser photocoagulation method. The vascular ligation method burns and ligates the central retinal vein to cause the retinal vein to flow back unsmoothly, and the vascular recanalization time is long, but the molding process is complex, the technical difficulty is high, the destructiveness is high, and infection easily occurs after operation to death of animals; the diathermy method can accurately position blood vessels, cause vasoconstriction and damage blood vessels through electric heating, and immediately block the blood vessels, but the method is characterized in that the blood vessels are blocked by utilizing electric heat energy to destroy vascular endothelium, different from human RVO forming mechanism, clinical guideline needs to be further researched, endothelin-1 is a powerful vasoconstrictor peptide, different in vascular reactions caused by injecting ET-1 with different concentrations into vitreous cavities, RVO caused by the ET-1 with proper concentrations is caused by serious vasospasm, only suitable for researching electrophysiological changes of retina ischemia monkeys, the laser photocoagulation retinal vein method is a noninvasive operation, laser is directly shot at the position of the blood vessels which are to be blocked by utilizing heat energy, the blocked blood vessels have higher recanalization rate and earlier recanalization time, repeated photocoagulation is needed for a plurality of times, serious thermal damage to retinal pigment epithelial cells and photoreceptor cells is easily caused by vitreous body bleeding caused by vascular wall damage, the combined fundus observation is realized by injecting a photosensitizing medicine vein injection, the photosensitizing medicine is carried out on the retinal vein in combination with a model of the cornea vein model, the model is formed by directly irradiating the laser vein model from the left eye to the left eye, the model and the cornea model is formed by the laser model before the cornea is irradiated from the left model to the right model, the model is formed by the cornea model, and the model is formed by the laser model is immediately before the model is formed by the cornea is formed, is easy to cause excessive damage to normal retinal tissues around blood vessels. Thus, it is highly necessary to develop animal models that have good reproducibility and that can exhibit pathological processes similar to those of RVO diseases for research in etiology, pathology and therapeutics. Disclosure of Invention The invention aims to provide a construction method of a retinal vein occlusion animal model. The invention provides a construction method of a retinal vein occlusion animal model, which comprises the following steps of firstly carrying out surface anesthesia on an eye of an anesthetized non-human primate, then sterilizing eye skin, eyelid margin and conjunctival sac, then establishing two scleral tunnels for eyeball puncture, respectively inserting an illumination optical fiber and an intraocular laser optical fiber into the two scleral tunnels, then injecting a photosensitizer through limb vein, and immediately carrying out laser photocoagulation on retinal vein under illumination of the illumination optical fiber after injection, thus obtaining the retinal vein occlusion animal model. Further, the inject