CN117256554A - C57BL/6J mouse neovascular eye disease model and construction method thereof - Google Patents
C57BL/6J mouse neovascular eye disease model and construction method thereof Download PDFInfo
- Publication number
- CN117256554A CN117256554A CN202311162238.XA CN202311162238A CN117256554A CN 117256554 A CN117256554 A CN 117256554A CN 202311162238 A CN202311162238 A CN 202311162238A CN 117256554 A CN117256554 A CN 117256554A
- Authority
- CN
- China
- Prior art keywords
- model
- mouse
- neovascular
- argon laser
- retinal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 208000030533 eye disease Diseases 0.000 title claims abstract description 14
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 title claims abstract description 12
- 238000010276 construction Methods 0.000 title abstract description 6
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims abstract description 34
- 229910052786 argon Inorganic materials 0.000 claims abstract description 17
- 210000001775 bruch membrane Anatomy 0.000 claims abstract description 13
- 210000001957 retinal vein Anatomy 0.000 claims abstract description 10
- 230000001678 irradiating effect Effects 0.000 claims abstract description 8
- 210000003583 retinal pigment epithelium Anatomy 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims description 12
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 10
- 230000003902 lesion Effects 0.000 claims description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 241000699670 Mus sp. Species 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 5
- 229940079593 drug Drugs 0.000 claims description 3
- 206010002091 Anaesthesia Diseases 0.000 claims description 2
- 230000037005 anaesthesia Effects 0.000 claims description 2
- 230000015556 catabolic process Effects 0.000 claims description 2
- 230000002911 mydriatic effect Effects 0.000 claims description 2
- 238000012216 screening Methods 0.000 claims description 2
- FKENQMMABCRJMK-RITPCOANSA-N sulbactam Chemical compound O=S1(=O)C(C)(C)[C@H](C(O)=O)N2C(=O)C[C@H]21 FKENQMMABCRJMK-RITPCOANSA-N 0.000 claims description 2
- 229960005256 sulbactam Drugs 0.000 claims description 2
- 208000005590 Choroidal Neovascularization Diseases 0.000 abstract description 34
- 206010060823 Choroidal neovascularisation Diseases 0.000 abstract description 34
- 210000001525 retina Anatomy 0.000 abstract description 9
- 206010029113 Neovascularisation Diseases 0.000 abstract description 7
- 208000022873 Ocular disease Diseases 0.000 abstract 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 9
- 230000006378 damage Effects 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 8
- 238000012014 optical coherence tomography Methods 0.000 description 8
- 230000002207 retinal effect Effects 0.000 description 8
- 208000007135 Retinal Neovascularization Diseases 0.000 description 6
- 210000003161 choroid Anatomy 0.000 description 6
- 238000010171 animal model Methods 0.000 description 5
- 210000004204 blood vessel Anatomy 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 4
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000002792 vascular Effects 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- 206010012689 Diabetic retinopathy Diseases 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000000465 moulding Methods 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 201000004569 Blindness Diseases 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- 201000006165 Kuhnt-Junius degeneration Diseases 0.000 description 2
- 206010038933 Retinopathy of prematurity Diseases 0.000 description 2
- 208000000208 Wet Macular Degeneration Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 206010064930 age-related macular degeneration Diseases 0.000 description 2
- 238000002583 angiography Methods 0.000 description 2
- 210000004087 cornea Anatomy 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- NJDNXYGOVLYJHP-UHFFFAOYSA-L disodium;2-(3-oxido-6-oxoxanthen-9-yl)benzoate Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=CC(=O)C=C2OC2=CC([O-])=CC=C21 NJDNXYGOVLYJHP-UHFFFAOYSA-L 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 208000004644 retinal vein occlusion Diseases 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 101100188551 Arabidopsis thaliana OCT2 gene Proteins 0.000 description 1
- 206010055665 Corneal neovascularisation Diseases 0.000 description 1
- 101000742579 Homo sapiens Vascular endothelial growth factor B Proteins 0.000 description 1
- 101000742596 Homo sapiens Vascular endothelial growth factor C Proteins 0.000 description 1
- 101000742599 Homo sapiens Vascular endothelial growth factor D Proteins 0.000 description 1
- 208000010415 Low Vision Diseases 0.000 description 1
- 208000001344 Macular Edema Diseases 0.000 description 1
- 206010025415 Macular oedema Diseases 0.000 description 1
- 241000218378 Magnolia Species 0.000 description 1
- 208000006550 Mydriasis Diseases 0.000 description 1
- XKLMZUWKNUAPSZ-UHFFFAOYSA-N N-(2,6-dimethylphenyl)-2-{4-[2-hydroxy-3-(2-methoxyphenoxy)propyl]piperazin-1-yl}acetamide Chemical compound COC1=CC=CC=C1OCC(O)CN1CCN(CC(=O)NC=2C(=CC=CC=2C)C)CC1 XKLMZUWKNUAPSZ-UHFFFAOYSA-N 0.000 description 1
- 206010052143 Ocular discomfort Diseases 0.000 description 1
- 101150062285 PGF gene Proteins 0.000 description 1
- 102100035194 Placenta growth factor Human genes 0.000 description 1
- 208000017442 Retinal disease Diseases 0.000 description 1
- 206010038923 Retinopathy Diseases 0.000 description 1
- 102100038217 Vascular endothelial growth factor B Human genes 0.000 description 1
- 102100038232 Vascular endothelial growth factor C Human genes 0.000 description 1
- 102100038234 Vascular endothelial growth factor D Human genes 0.000 description 1
- 208000035307 Vitreous adhesions Diseases 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- -1 compound topiramate Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 201000000159 corneal neovascularization Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 208000002780 macular degeneration Diseases 0.000 description 1
- 201000010230 macular retinal edema Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 229920002492 poly(sulfone) Polymers 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229960000213 ranolazine Drugs 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000004263 retinal angiogenesis Effects 0.000 description 1
- 230000037390 scarring Effects 0.000 description 1
- 210000003786 sclera Anatomy 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229960004394 topiramate Drugs 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 230000004382 visual function Effects 0.000 description 1
- 210000004127 vitreous body Anatomy 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Endocrinology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Diabetes (AREA)
- Animal Husbandry (AREA)
- Gastroenterology & Hepatology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Rheumatology (AREA)
- Toxicology (AREA)
- Urology & Nephrology (AREA)
- Epidemiology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention provides a C57BL/6J mouse neovascular eye disease model and a construction method thereof. The construction method includes irradiating a mouse Bruch's membrane and retinal pigment epithelium covered thereon with an argon laser, and irradiating a retinal vein trunk with an argon laser fundus. Compared with a Choroidal Neovascularization (CNV) model, the retina+choroidal neovascularization (CNV+RNV) model constructed by the invention generates more durable and stable neovascularization, and is suitable for being used as a C57BL/6J mouse neovascularization type ocular disease model.
Description
Technical Field
The invention belongs to the field of animal models, and particularly relates to a C57BL/6J mouse neovascular eye disease model and a construction method thereof.
Background
The neovascular eye disease refers to eye diseases affecting vision caused by pathological changes such as bleeding, exudation, hyperplasia and the like due to the growth of new blood vessels, and is one of the most serious blindness eye diseases worldwide. Including Diabetic Retinopathy (DR), neovascular age-related macular degeneration (nAMD), retinal Vein Occlusion (RVO), retinopathy of prematurity (ROP), choroidal neovascularization (Choroidal Neovascularition, CNV).
The current animal models of neovascular eye diseases are mainly a retinal neovascular (Retinal Neovascularization, RNV) model, a choroidal neovascular model, and a corneal neovascular model. The most commonly used retinal neovascular model is an oxygen-induced retinopathy (OIR) model, which has the advantages that neovascular is easy to induce, the neovascular area is easy to quantitatively analyze, and the defects that the high oxygen volume fraction adopted by most OIR animal models is not completely consistent with clinical application and the model duration is short.
The laser-induced CNV model is one of the most widely used models, which is relatively fast to build and has good reproducibility, and has the disadvantages that the technical differences of each laser application cause lesions to often show large differences in size between animals even in the same 1 eye, and have poor stability in small animals. The cornea neovascular model mainly comprises an alkali injury model, a suture injury model and a cornea microcapsule model, and has less overall application.
The choroid is located between the retina and sclera, above which is spread the blood vessels of the michelia, providing oxygen and nutrition to the outer layers of the retina. Under various complications, abnormal new blood vessels are produced in the choroid and invade under the retina, causing subretinal exudation, hemorrhage, and scarring. And, the macular edema becomes thicker in the macular area accumulated on the retina, causing permanent damage to visual function. Retinal neovascularization originates from the disc surface and the small veins of the retina, grows along the retinal surface, can grow into the vitreous body at sites where there are vitreous adhesions, and contains varying amounts of fibrous tissue, known as neovascular membranes. Most are caused by retinal large area capillary occlusion and chronic ischemia, i.e., associated with the production and release of vascular endothelial growth factor. Patients may be associated with ocular discomfort and severe patients may also experience symptoms such as reduced vision or blindness.
Disclosure of Invention
The invention aims to overcome the defect that a novel vascular animal model which is simple and stable to operate and long in service life is lacked in the prior art, and provides a C57BL/6J mouse novel vascular eye disease model and a construction method thereof.
The invention solves the technical problems through the following technical proposal.
A first aspect of the present invention provides a method of constructing a model of ocular neovascular disease in a mouse, the method comprising irradiating a Bruch's membrane and retinal pigment epithelium overlying the Bruch's membrane with an argon laser and irradiating retinal vein stems with an argon laser fundus.
In some embodiments of the invention, the irradiation of retinal vein stems with an argon laser fundus is performed after irradiation of the mouse Bruch's membrane and overlying retinal pigment epithelium with an argon laser.
In some embodiments of the invention, the argon laser has an emission wavelength of 532nm and an irradiation condition of 190mW for 0.1s.
In the invention, the irradiation adopts a fixed laser to irradiate the laser fundus.
In some embodiments of the invention, the method comprises: fundus irradiation with argon laser and retinal pigment epithelium covering it, breakdown of the Bruch membrane, creating 2 lesion sites of 75 μm diameter on the Bruch membrane; also included was irradiation of the retinal vein stem with an argon laser fundus, producing 2 lesion sites of 75 μm diameter on the retinal vein stem.
In some embodiments of the invention, the irradiation of retinal vein stems with an argon laser fundus occurs after creating a lesion on the Bruch's membrane.
According to the above embodiment, a model of mouse choroid and retinal neovascularization (CNV+RNV) can be constructed.
In some embodiments of the invention, the mice are anesthetized and mydriatic prior to irradiation.
In some preferred embodiments of the invention, the anesthesia employs sultai and selazine.
In some embodiments of the invention, the dosage of the sulbactam is 80mg/kg and the dosage of the selazine is 5mg/kg.
In some embodiments of the invention, the mouse is a C57BL/6J mouse.
In a second aspect, the present invention provides a model of ocular neovascular diseases in mice constructed by the method according to the first aspect.
In some embodiments of the invention, the model is a choroidal+retinal neovascularization (cnv+rnv) model.
In a third aspect, the present invention provides the use of a mouse neovascular eye disease model according to the second aspect for screening a medicament for preventing and/or treating neovascular eye disease.
On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that: according to the invention, through a laser-induced Choroidal Neovascularization (CNV) model and a choroidal and retina neovascularization (CNV+RNV) model, ophthalmic examination and molecular biological experiments are carried out on the two neovascularization models, and experimental results show that compared with the choroidal neovascularization model, the choroidal and retina neovascularization model has a more stable and durable effect, and is more suitable for being used as a model for researching ophthalmic neovascularization diseases and related medicines.
Drawings
Fig. 1 is a schematic representation of Optical Coherence Tomography (OCT) results from day 7 (D7) and day 30 (D30) following choroidal neovascularization model laser.
Fig. 2 is a schematic representation of Optical Coherence Tomography (OCT) results from day 7 (D7) and day 30 (D30) following choroidal + retinal neovascular model laser.
Fig. 3 is a sodium Fluorescein Fundus Angiography (FFA) performed on days 7 (D7) and 30 (D30) after choroidal neovascularization model laser.
Fig. 4 is a sodium Fluorescein Fundus Angiography (FFA) performed on days 7 (D7) and 30 (D30) after choroidal+retinal neovascular model laser.
FIG. 5 shows D30 Vascular Endothelial Growth Factor A (VEGFA) protein expression following laser irradiation of a choroidal neovascularization model and a choroidal+retinal neovascularization model.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
Examples in the present invention, the effect of the choroidal+ retinal neovascular model was more stable and durable than that of the choroidal neovascular model
1. Test materials
1.1VEGFA antibodies
VEGF is an important group of mediators that stimulate neovascularization and increase the permeability of existing blood vessels, playing a key role in normal development and wound healing. VEGF includes the types VEGFA, VEGFB, VEGFC, VEGFD, VEGFE and PLGF, and VEGFA is most commonly found in drug studies in relation to vascular endothelial cell proliferation, migration, and microvascular formation. When VEGFA is overexpressed in the eye, abnormal vascular proliferation, vascular leakage, etc. can result, which in turn, causes various ophthalmic diseases such as macular degeneration, diabetic retinopathy, corneal neovascularization, etc. Medicaments with VEGF (mainly VEGFA) as a target point open a new way for treating ocular neovascular diseases.
1.2 test animals
18C 57BL/6J mice were male. Purchased from beijing vernalia laboratory animal technology limited. Polysulfone mouse box with laying padding and nesting materials, temperature: 20-26 ℃, humidity: 40% -70%, and the adaptation time is more than 3 days before the experiment.
2. Test method
2.1 packet case
Animals were divided into 2 test groups, choroidal neovascularization model (CNV) and choroidal+retinal neovascularization model (cnv+rnv), respectively. The grouping is detailed in Table 1.
Table 1: grouping situation
| Grouping | Quantity of | Test |
| CNV | 9 | FFA、OCT、Western blot |
| CNV+RNV | 9 | FFA、OCT、Western blot |
2.2 test methods
After anaesthetizing a C57BL/6J mouse with 80mg/kg of sultai+5 mg/kg of ranolazine, a compound topiramate eye drop (participating in the world, pharmaceutical Co., ltd.) is used for mydriasis, a fixed laser is used for fundus irradiation with argon laser (190 mW,0.1 s) to break down the Bruch membrane and the retinal pigment epithelium covered on the Bruch membrane, and 4 damage points with the diameter of 75 mu m are burned out, thus obtaining the choroidal neovascularization model of the C57 mouse. After the choroidal neovascularization model is molded (2 damage points), the retinal vein trunk is irradiated by the fundus again by argon laser (190 mW,0.1 s), 2 damage points with the diameter of 75 mu m are burned out, and the laser is accurately focused on the vein when irradiating the vein, so that adjacent arteries are prevented from being damaged. The choroid and retina neovascular model is obtained.
After the molding, the ophthalmic OCT examination was performed on the 7 th day (D7) and the 30 th day (D30) to determine the position and condition of the fluorescent spot.
Ophthalmic FFA examinations were performed on day 7 (D7) and day 30 (D30) after molding was completed. Observing FFA conditions in each period, calculating the number and range of strong fluorescence spots generated by fluorescein leakage 8-10 minutes after the shadow making so as to evaluate pathological changes of choroid and retinal angiogenesis, and carrying out statistical treatment; and grading the lesion degree.
Mice were euthanized at day 30 (D30) after molding was completed, retinal choroid was harvested, and Western blot was performed.
During the post-laser observation period, the general status of the animals was observed 1 time a day, and the general status of the animals was recorded.
3. Test results
3.1 general case
The animals were active spontaneously during the observation period, were normal in diet, were well-conditioned, and no obvious abnormal response was observed.
3.2OCT case
The location and recovery of the plaques in the OCT results are consistent with the FFA results, see in particular figures 1 and 2.
3.3 laser induced CNV model and CNV+RNV model in number of fluorescent spots different.
After the C57BL/6J mice are subjected to laser modeling, FFA results show that: after laser, D30, compared with the CNV model, the number of the laser damage spots with fluorescence leakage of the CNV+RNV model group is obviously increased, and the fluorescence leakage of the laser damage spots is stronger than that of the CNV. The results are detailed in Table 2. See in particular fig. 3 and 4.
Table 2: number of laser damage spots leaking fluorescent spots after laser modeling of C57BL/6J mice
3.4Western blot results showed that the CNV+RNV group highly expressed VEGFA compared to the CNV model group (FIG. 5).
3.5 conclusion
Under the laboratory condition, after the C57BL/6J mice are subjected to laser modeling, FFA results show that: compared with the CNV model, the fluorescence leakage of the CNV+RNV model group is obviously increased, and the fluorescence leakage of the laser lesion part is stronger than that of the CNV. Western blot results also show that the expression level of the VEGFA protein in the CNV+RNV model group is higher. The determination result shows that the CNV+RNV model has more stable and durable effect of generating new blood vessels, and is more suitable for being used as an ophthalmic neovascular type eye disease model.
Claims (7)
1. A method of constructing a model of ocular neovascular disorder in a mouse, the method comprising irradiating a Bruch's membrane and retinal pigment epithelium overlying the Bruch's membrane with an argon laser and irradiating retinal vein stems with an argon laser fundus.
2. The method of claim 1, wherein the argon laser has an emission wavelength of 532nm and an irradiation condition of 190mw for 0.1s.
3. The method according to claim 2, wherein the method comprises: fundus irradiation with argon laser and retinal pigment epithelium covering it, breakdown of the Bruch membrane, creating 2 lesion sites of 75 μm diameter on the Bruch membrane; and irradiating the retinal vein stem with an argon laser fundus, creating 2 lesion spots of 75 μm diameter on the retinal vein stem.
4. A method according to any one of claims 1 to 3, wherein the mice are anesthetized and mydriatic prior to irradiation; preferably, the anesthesia adopts sultai and cerazine;
more preferably, the dosage of the sulbactam is 80mg/kg, and the dosage of the selazine is 5mg/kg.
5. The method of any one of claims 1-4, wherein the mouse is a C57BL/6J mouse.
6. A model of ocular neovascular disease in mice constructed by the method of any one of claims 1-5.
7. Use of the mouse neovascular eye disease model of claim 6 for screening a drug for preventing and/or treating neovascular eye disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311162238.XA CN117256554A (en) | 2023-09-08 | 2023-09-08 | C57BL/6J mouse neovascular eye disease model and construction method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311162238.XA CN117256554A (en) | 2023-09-08 | 2023-09-08 | C57BL/6J mouse neovascular eye disease model and construction method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117256554A true CN117256554A (en) | 2023-12-22 |
Family
ID=89220642
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311162238.XA Pending CN117256554A (en) | 2023-09-08 | 2023-09-08 | C57BL/6J mouse neovascular eye disease model and construction method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117256554A (en) |
-
2023
- 2023-09-08 CN CN202311162238.XA patent/CN117256554A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Al-Aqaba et al. | Corneal nerves in health and disease | |
| Bandello et al. | Pathophysiology and treatment of diabetic retinopathy | |
| Ryan | The development of an experimental model of subretinal neovascularization in disciform macular degeneration. | |
| Aihara et al. | Experimental mouse ocular hypertension: establishment of the model | |
| Provis et al. | Astrocytes and blood vessels define the foveal rim during primate retinal development | |
| Ebneter et al. | Investigation of retinal morphology alterations using spectral domain optical coherence tomography in a mouse model of retinal branch and central retinal vein occlusion | |
| Khayat et al. | Animal models of retinal vein occlusion | |
| TW200302226A (en) | Methods for treating ocular neovascular diseases | |
| Figus et al. | The supraciliary space as a suitable pathway for glaucoma surgery: Ho-hum or home run? | |
| Gallego-Ortega et al. | Functional and morphological alterations in a glaucoma model of acute ocular hypertension | |
| Oatts et al. | In vitro and in vivo comparison of two suprachoroidal shunts | |
| Irvine et al. | Radiation retinopathy: an experimental model for the ischemic--proliferative retinopathies. | |
| Semkova et al. | Investigation of laser-induced choroidal neovascularization in the rat | |
| Augustin | Intravitreal Steroids | |
| Sørensen | Subretinal surgery: functional and histological consequences of entry into the subretinal space | |
| Zhang et al. | Experimental models and examination methods of retinal detachment | |
| Kumar et al. | A robust microbead occlusion model of glaucoma for the common marmoset | |
| WALLOW et al. | Focal photocoagulation of diabetic macular edema: a clinicopathologic case report | |
| CN113273546A (en) | Application of lauromacrogol in preparation of chronic ocular hypertension animal model and animal model | |
| Ejstrup et al. | Electrophysiological consequences of experimental branch retinal vein occlusion in pigs and the effect of dorzolamide | |
| CN117256554A (en) | C57BL/6J mouse neovascular eye disease model and construction method thereof | |
| CN111226865A (en) | Method for constructing tree shrew choroidal neovascularization animal model | |
| Reiner et al. | Neural control of ocular blood flow | |
| Miller et al. | A single intravitreal injection of ranibizumab provides no neuroprotection in a nonhuman primate model of moderate-to-severe nonarteritic anterior ischemic optic neuropathy | |
| CN112568187B (en) | Method for constructing novel mouse model with retinal vein occlusion |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |