A kind of method for building up of chronic intraocular hypertension animal modelTechnical field
The present invention relates to medical animal model field, be specifically related to the method for building up of a kind of chronic intraocular hypertension animal model.
Background technology
Glaucoma is a kind of irreversibility optic nerve degeneration, for whole world second diseases causing blindness.The pathogenesis of its complexity is the most still not very clear, although Bulbi hypertonia is not the single factor that glaucoma regards infringement, but Bulbi hypertonia is one of factor the most dangerous in Deterioration of Optic Nerve in Glaucoma generation and development mechanism.In GLAUCOMA RESEARCH field, animal model has become important research tool.Glaucoma animal model has important effect for the damage mechanisms and remedy measures being better understood from glaucomatous optic neuropathy.Bulbi hypertonia is the topmost risk factor of Deterioration of Optic Nerve in Glaucoma, so the animal model of most of GLAUCOMA RESEARCH remains induced animal Bulbi hypertonia, and then studies the aspects such as the physiology of glaucoma, pathology, pharmacology.Preferably experimental animal model requires: its construction method is simple, with low cost, it is easy to raise and control, and the intraocular pressure of rising remains stable, and the distinctive pathological change of retina optic nerve occurs.
The animal being currently used for making glaucoma model is low mammal such as grade, mainly has rabbit, Mus, monkey.Although Mus is prone to transgenic technology transformation and breeding, and the structure of rathole is similar with human eye at a lot of aspects, and such as girder, Schlemm pipe, corpus ciliare, retinal vessel etc., but rathole is relatively small, is unfavorable for being operated.The Ocular hypertensive model of the monkey of induced with laser is the optimal Bulbi hypertonia animal model generally acknowledged at present, but because involving great expense, operates more complicated, and domestic application is few.Although lagophthalmos has Morphological Differences with the chamber-angle structure of human eye, but ah outflow passage is identical, has continuous print endotheliocyte liner.And rabbit price is inexpensive, personality is docile, be easily obtained and operate, therefore rabbit is most commonly used for making the laboratory animal of glaucoma model.
It is divided into according to Bulbi hypertonia Forming Mechanism and medicine site of action and acts on angle of anterior chamber (and girder) type, such as injected into anterior chambers hyaluronic acid, methylcellulose, hemocyte, Carbomer liquid, back room injection 5. Chymotrypsin, laser photocoagulation trabecular reticulum, the method such as eyes external corticosteroid hormone；Act on the most conventional scleral surface intravenous injection hypertonic saline method (Morre-Morrison model) of sclera aqueous humor drains intravenous type, burn episcleral aqueous humor drainage vein method (Shareef-Sharma model) and modification method；Additionally, also have the spontaneous mouse of some special germlines and transgenic technology to build glaucoma model, such as the mouse inbred lines of DBA/2J and AKXD-28/Ty, utilize the natural glaucoma model etc. that gene targeting builds.The most various models pathologic in terms of retinal ganglial cells (RGCs), optic nerve, retinal function is changed and be evaluated, assess the Positive and Negative Aspects of various model, preferably build, in chronic intraocular hypertension glaucoma Mechanism Study, the modeling animal agreed with mutually with clinic and modeling method provides foundation, the more mechanism of in-depth study chronic intraocular hypertension glaucoma, provides new thinking for its treatment.
Although the method making Bulbi hypertonia animal model has a lot, but current glaucoma animal model great majority are all acute high IOP models, although intraocular pressure raises, but its Bulbi hypertonia is held time short, and it is big to fluctuate, it is difficult to simulate the characteristic optic nerve lesion of glaucoma.
Summary of the invention
In order to overcome the defect of prior art, the invention provides the method for building up of a kind of chronic intraocular hypertension animal model.
The technical solution that the present invention uses is: the method for building up of a kind of chronic intraocular hypertension animal model, comprise the following steps: animal pattern bondage is lain on the back in operating-table by (1), art eye sterile drape, put eye speculum, rinse conjunctival sac, under art eye conjunctiva after the anesthesia of local profit, do superior rectus traction sutures and fix；
(2) timepiece dial telegoniometer is pressed, in 11 positions to 1 position edge ring bulbar conjunctiva incision, centered by 12 positions, place makees the shallow-layer scleral flap of 1/3 scleral thickness of size 4*4mm with limbus of corneae as substrate, size 3*3mm is done again below shallow-layer scleral flap, the Deep Layer of Sclera lobe of thickness 2/3 scleral thickness, and peel off to cornea direction, find Schlemm to manage and cut Schlemm pipe outer wall；
(3) in Schlemm pipe two ends, polymer hyaluronic acid is injected, fiber duct conduit is passed through opening, in 9 or 3 o ' clock orientation limbus of corneae at 1.0 mm, flat iris direction row paracentesis of anterior chamber, light pressure puncture orifice trailing edge, release water and reduce intraocular pressure, anterior chamber is reduced pressure and fiber duct is inserted after puncturing the broken ends of fractured bone of Schlemm pipe, 360 degree or 270 degree or 180 degree or 90 degree are detoured along Schlemm pipe, cut off conduit, it is placed in Schlemm blocking Schlemm pipe, stop ah outflow, sew up scleral flap, , paracentesis of anterior chamber mouth injection balance saline solution, sew up conjunctival flap, give anti-inflammatory drug, finally set up the animal model that intraocular pressure raises.
Described step (1) use dilution iodophor solution rinse conjunctival sac.
Described step (1) use the lignocaine of 0.4ml 2% carry out profit anesthesia in local under art eye conjunctiva.
Described step (1) use 30G syringe needle inject polymer hyaluronic acid in Schlemm pipe two ends.
In described step (3), fiber duct includes a cylindrical solid conduit, and described tube at one end is spherical structure.
The long 10cm of described fiber duct, diameter 200 μm, the spherical diameter of described tube at one end is 250 μm.
Described fiber duct uses polypropylene material to make.
The invention has the beneficial effects as follows: the invention provides the method for building up of a kind of chronic intraocular hypertension animal model, place material by operation and block its aqueous humor flow pass, intraocular pressure is caused to raise the chronic glaucoma animal model of optic nerve lesion, the chronic intraocular hypertension animal model produced by this method, its intraocular pressure can regulate and control according to the scope difference of blocking Schlemm.And the fiber duct of different specification size can be selected according to different animals kind Schlemm caliber, complete Schlemm blockage, thus produce the ocular hypertension model of many animals type.In chronic intraocular hypertension glaucoma Mechanism Study, preferably build the modeling animal agreed with mutually with clinic and modeling method, provide basis for carrying out chronic glaucoma optic nerve injury mechanism.
Accompanying drawing explanation
Fig. 1 is fiber duct structural representation of the present invention.
Detailed description of the invention
In conjunction with Fig. 1, the present invention is further described: operating procedure: operation needs a long 10cm diameter 200 μm solid fibers conduit being made by polypropylene material, the a diameter of 250 μm local bulkiness of conduit head end spherical, damages Schlemm tube wall tissue when preventing from intubating.
Animal pattern extremity bondage is lain on the back in operating-table, art eye routine disinfection drape, put eye speculum, dilution iodophor solution rinses conjunctival sac, take after locally moistening anesthesia under 0.4ml 2% lignocaine Rhizoma Atractylodis Macrocephalae eye conjunctiva, do superior rectus traction sutures to fix, by timepiece dial telegoniometer, in 11 positions to 1 position edge ring bulbar conjunctiva incision, centered by 12 positions, place makees the shallow-layer scleral flap of 1/3 scleral thickness of the size about 4*4mm with limbus of corneae as substrate, a size about 3*3mm, the Deep Layer of Sclera lobe of thickness about 2/3 scleral thickness is done again below shallow-layer scleral flap.And peel off to cornea direction, find Schlemm to manage and cut Schlemm pipe outer wall, in Schlemm pipe two ends, injecting polymer hyaluronic acid (Healon GV) with 30G syringe needle, enable the duct to easily by opening.In 9 or 3 o ' clock orientation limbus of corneae at 1.0 mm, flat iris direction row paracentesis of anterior chamber, light pressure puncture orifice trailing edge, releases appropriate aqueous humor and reduces intraocular pressure, and anterior chamber is reduced pressure and microtubular is inserted after puncturing the broken ends of fractured bone of Schlemm pipe, difference according to required target intraocular pressure, detour 360 degree or 270 degree or 180 degree or 90 degree along Schlemm pipe respectively, cut off conduit, be placed in Schlemm blocking Schlemm pipe, stop ah outflow, cause the model that its intraocular pressure raises.Finally with 10-0 polypropylene line tight suture scleral flap 4-6 pin, paracentesis of anterior chamber mouth injection balance saline solution is observed, and sews up with the non-leakage watertight that is considered as, then with 10-0 polypropylene suturing with thread management conjunctival flap 2 pin.The postoperative anti-inflammatory drug that gives, monitors intraocular pressure, prosthomere response situation.
The present invention places material by operation and blocks its aqueous humor flow pass, intraocular pressure is caused to raise the glaucoma animal model of optic nerve lesion, the chronic intraocular hypertension animal model produced by this method, its intraocular pressure can regulate and control according to the scope difference of blocking Schlemm.And the fiber duct of different specification size can be selected according to different animals kind Schlemm caliber, complete Schlemm blockage, thus produce the ocular hypertension model of many animals type..The fiber duct specification that lagophthalmos, the animal model of monkey eye use can use long 10cm diameter 200 μm, and conduit head end spherical diameter is 250 μm, the optional conduit specification matched with its Schlemm caliber of other animals.In chronic intraocular hypertension glaucoma Mechanism Study, preferably build the modeling animal agreed with mutually with clinic and modeling method, provide basis for carrying out chronic glaucoma optic nerve injury mechanism.
The above is only the preferred embodiment of the present invention, and protection scope of the present invention is not limited merely to above-described embodiment, and all technical schemes belonged under thinking of the present invention belong to protection scope of the present invention.It should be pointed out that, for those skilled in the art, some improvements and modifications without departing from the principles of the present invention, these improvements and modifications also should be regarded as protection scope of the present invention.