CN102198278A - Rhesus monkey retinal degeneration model and drug screening method thereof - Google Patents
Rhesus monkey retinal degeneration model and drug screening method thereof Download PDFInfo
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- CN102198278A CN102198278A CN2010101311480A CN201010131148A CN102198278A CN 102198278 A CN102198278 A CN 102198278A CN 2010101311480 A CN2010101311480 A CN 2010101311480A CN 201010131148 A CN201010131148 A CN 201010131148A CN 102198278 A CN102198278 A CN 102198278A
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Abstract
The invention provides a preparation method of a rhesus monkey retinal degeneration model, which is to apply sodium iodate with the dose of 15-75mg/kg to a rhesus monkey. The invention also provides a method for screening the medicine for treating the retinal degeneration. The invention firstly tries to inject the sodium iodate into the rhesus monkey body by intravenous injection to construct a primate retinal degeneration model, screens a proper modeling dose and establishes a systematic model evaluation system.
Description
Technical field
The present invention relates to a kind of Rhesus Macacus retinal degeneration animal model, particularly, is that sodium iodate is applied to the animal model that Rhesus Macacus is prepared from.
Background technology
Retinal degeneration is one of main oculopathy of current blinding, and the foundation of stable animal model is the key that solves related scientific research and clinical problem.The model of current retinal degeneration is primarily aimed at rodent, rabbit etc., as: wangkai, etc., sodium iodate is induced the research of rabbit retina and optic nerve transneuronal degeneration, "
Ophthalmology research", 2008 26 7 phases of volume, can induce the retina of laboratory animal and optic nerve transneuronal degeneration to occur by the intravenous injection of experimental observation sodium iodate.Concrete grammar is: 48 livid purple blue rabbits, 8 for matched group. and remain 40 carries out sodium iodate intravenous injection (40mg/kg), observes injection afterwards 1 week, 2 weeks, the pathological change of 1,4,18 month laboratory animal retina and optic nerve, and compare with matched group.Adopt DNA terminal transferase mediated dUTP nick end labeling breach end-labelling (TUNEL) labelling apoptotic cell; Chromotropic acid 2R dyes the optic nerve myelin; Guillery Shirra-Webster method is dyed the nerve fiber of degeneration; The neuron of inner nuclear layer of retina is at sodium iodate injection back 1 week beginning apoptosis as a result; Retinal ganglial cells (RGCs) is at injection beginning in back 1 month apoptosis; After injection 4 months and 18 months, visible optic nerve demyelination and degeneration in various degree; The intravenous injection of conclusion sodium iodate can be induced laboratory animal retina transneuronal degeneration.
Chen Qin, epidermal growth factor treatment sodium iodate is induced retinal pigment epithelium degeneration scanning electron microscopic observation, "
Ophthalmology research", 1996 14 4 phases of volume, reported to rabbit vein and injected sodium sulfonate, injected once again in 48 hours and 72 hours respectively at interval, occurred part eye, scanning electron microscopic observation RPE cell appearance and quantity on the 7th day, inject the hEGF extracting solution in the surplus right eye ball vitreous body, left eye is annotated the equivalent normal saline.
Inhuman primate is very approaching with the mankind on kind, does not still have the report of the model of primates retinal degeneration at present.
Summary of the invention
The invention provides a kind of Rhesus Macacus retinal degeneration Preparation of model method, the present invention also provides a kind of method of screening the medicine of treatment retinal degeneration.
The invention provides a kind of Rhesus Macacus retinal degeneration Preparation of model method, is that the sodium iodate of 15-75mg/kg is applied to Rhesus Macacus with dosage.Further preferably, the dosage of described sodium iodate is 50mg/kg.
The present invention also provides the purposes of animal model in the medicine of screening treatment retinal degeneration of described method preparation.
The present invention also provides a kind of method of screening the medicine of treatment retinal degeneration, and it comprises the steps:
A, be applied to Rhesus Macacus, set up Rhesus Macacus retinal degeneration model with sodium iodate;
B, drug candidate is applied to the degeneration model;
C, observe the influence situation of drug candidate, and mark, estimates potential at the medicine for the treatment of retinal degeneration to the quantizating index of retinal degeneration, degree of injury.
Wherein, the application dosage of the sodium iodate described in the step a is: 15-75mg/kg.
Wherein, the application dosage of the sodium iodate described in the step a is: 50mg/kg.
The present invention attempts the sodium iodate intravenous injection is gone into Rhesus Macacus construct in vitro primates retinal degeneration model first, screens suitable modeling dosage, and sets up the model evaluation system of system.
Description of drawings
The amplitude change of flash of light ERG after Fig. 1 modeling
The change curve of the flash of light ERGa wave-wave width of cloth after Fig. 2 modeling
The change curve of the flash of light ERGb wave-wave width of cloth after Fig. 3 modeling
After the modeling of Fig. 4 50mg/kg group after the variation modeling of fundus photography 7 days optical fundus do not see obvious change, from 14 days, point-like, block pigmentation all appearred in the utmost point and periphery behind the retina, 28 days more obvious.
A large amount of transmitted fluorescence of representing the retinal pigment epithelium damage appearred in each one of retina after fluorescence fundus angiography changed modeling after the modeling of Fig. 5 50mg/kg group, oozed out on a small quantity.
HE coloration result A is normal monkey eyes retina after Fig. 6 modeling, pigment epithelium is level and smooth, the cell internuclear distance is close, each confluent monolayer cells etc. are big, marshalling, B is 3 week of a 30mg/kg group back retina, the pigment epithelium disappearance, the degeneration of cavity sample appears in the photoreceptor layer, but obvious change does not appear in inner each confluent monolayer cells.C is 6 week of a 50mg/kg group back retina, and pigment epithelium cell nuclear does not wait big, and internuclear distance difference is big, the degeneration of photoreceptor layer, and interior outer nuclear layer nucleus does not wait greatly, arranges and sees disorder.
The specific embodiment
The preparation of test example 1 animal model of the present invention
1, test objective
Set up the model that sodium iodate causes the Rhesus Macacus retinal degeneration, set up the model evaluation system of system.
2, test material
2.1 main agents
Anesthetis: 3% pentobarbital 30mg/kg, ketalar 50mg/ml
Atropine sulfate 10mg/2ml
Disinfectant: polyvidone povidone iodine 5.0g/L
Modeling agent: sodium iodate 50mg/ml
Optical fundus contrast agent: 20% fluorescein sodium, indole viridescent
2.2 key instrument, apparatus
The SL-2F slit lamp
Direct ophthalmoscope
JFY2EP-1000 eye electrophysiology instrument
Heidelberg fundus fluorescein angiography instrument
The Heidelberg fundus camera
2.3 laboratory animal
2.3.1 animal feeding management
Rhesus Macacus is raised in conventional animal room, Chinese medicine safety evaluatio center, national Chengdu, the preceding animal of experiment conforms and carries out the anthelmintic treatment, select healthy (female palpus unpregnancy) animal as animal subject, rustless steel monkey cage sub-cage rearing, the raising condition meets GB GB14925-2001,18 ± 26 ℃ of room temperatures, relative humidity 40 ± 70%.Clear and mischievous person's forage feed, tap water bottling are freely drunk, every day feed fruit.
Animal is hung on neck board and the cage and labels as animal identification marking.
2.3.2 laboratory animal screening
10 of regular grade adult healthy Rhesus Macacus, male and female half and half are intended breeding study base (production licence number: SCXK (river) 2004-013) by Chengdu, Sichuan safety animal and are introduced.Enter the room and carry out tuberculin, parasite, Salmonella, the Salmonella of congratulating inspection the last week.
Include standard in: 4-5 year, body weight 4-6kg, the healthy Rhesus Macacus of no B herpesvirus, monkey retrovirus, monkey leucovirus and immunodeficiency virus infection.Need through routine blood test after animal enters the room, go out the first screening that thrombotest, serum biochemistry are checked, get rid of great organ lesion.Ophthalmology illness is got rid of in the postsearch screening of splitting light-off, examination of ocular fundus, flash of light ERG inspection, fluorescein angiographic inspection afterwards.Twice normal animal of The selection result can be included this experimentation in.
3, test method
The method that adopts ulnar vein to inject is carried out the test of 15mg/kg, 30mg/kg, 50mg/kg, 4 dosage of 75mg/kg respectively to Rhesus Macacus, by amphiblestroid function damage after flash of light ERG, the fluorescein angiographic detection modeling; By splitting the damage of the inspection of turning off the light, fundus photography detection general form; Detect ultra micro Histological injury by fixedly HE dyeing of sclerous tissues, electron microscopic examination; Detect damage by blood blood routine blood test, blood biochemistry, the HE dyeing that goes out blood coagulation experiment and great internal organs to whole body organ function and form.Select suitable dosage according to comprehensive evaluation result.
4, result of the test
4.1 flash of light ERG: see Fig. 1, Fig. 2, Fig. 3
Analyze: significant change all appears in the wave amplitude of various dose group ERG after the modeling.The severe reaction of whole body appears in 75mg/kg group, and the ERG wave amplitude disappears, and occurs one after the modeling of 15mg/kg, 30mg/kg group and crosses the property reduction, return to modeling in 3 weeks before level.2 weeks of 50mg/kg dosage group reach minimum, and 3 weeks had recovery slightly, and 6 weeks steadily maintained reduced levels after modeling.
4.2 fundus photography (see figure 4)
4.3 fluorescence fundus angiography (see figure 5)
The (see figure 6) 4.4 retina HE dyes
Detect 4.5 great organopathy is of science
Sodium iodate (50mg/kg, 75mg/kg) vein gives Rhesus Macacus after June, observes its toxicity pathology reaction.
Microscopically is observed the regeneration of 75mg/kg group 1 animal (05648) renal cells, jejunum enteraden lamina propria fibrosis.Pathological changes occurs over just high dose group, can not get rid of the relevant diseases for drug-induced.
Microscopically is observed 75mg/kg group 1 animal (05648), 50mg/kg group 1 animal (06222) duodenal recess basilar part glandular epithelium vacuolar degeneration.Pathological changes all has generation in the high and low dose group, may be drug-induced.
Microscopically is observed the cystic dilatation of 50mg/kg group 1 animal (06222) colon enteraden, includes sphacelus, 1 animal (06111) atrophic gastritis.Pathological changes occurs in low dose group, can not get rid of the relevant diseases for drug-induced.
Spontaneous or sporadic pathological changes:
Microscopically is observed 75mg/kg group 1 animal (05648), the little granuloma of 50mg/kg group 1 animal (06111) liver.Due to illness the range degree is light, and the incidence rate that animal should disease is higher, the spontaneous pathological changes of polyphyly animal.
To sum up tell, sodium iodate (50mg/kg, 75mg/kg) vein gives Rhesus Macacus after June, and according to pathological examination results, two dosage digestive tract pathological changes polyphyly medicine all occurs and are correlated with, 50mg/kg is slighter, and 75mg/kg the change of kidney occurs and can not get rid of relevant with medicine.
5, discuss
ERG sees by flash of light, and 75mg/kg group wave amplitude disappears, can not quantitative statistics.One property crossed reduction appears after the modeling of 15mg/kg, 30mg/kg group, level before returning to modeling in 3 weeks, the proof function damage is also not serious, and the painted result of HE shows, the pigment epithelium disappearance only occurs, the degeneration of cavity sample appears in the photoreceptor layer, obvious change does not appear in inner each confluent monolayer cells, and we infer that this dosage has early stage acute injury to retina, but the compensatory capacity that does not exceed Rhesus Macacus, damage can recover.50mg/kg dosage group ERG 2 weeks of wave amplitude reach minimum, and 3 weeks had recovery slightly, and 6 weeks steadily maintained reduced levels after modeling, and the stage of stable development of long period is arranged, and were convenient to quantitative statistics.And HE is except that retinal pigment epithelium and the damage of photoreceptor layer, and interior outer nuclear layer nucleus does not wait greatly, and arrangement disorder has been verified the damage of sodium iodate to each layer of retina from the histology.Pigmentation appears in each one of fundus photography retina, and FFA shows that the characteristic window sample of blocked fluorescence and pigment epithelium damage is damaged, has verified the success of modelling from clinical angle.
6, sum up
50mg/kg sodium iodate intravenous injection can cause the Rhesus Macacus retinal degeneration, and degree of injury is moderate, and is little to other organ injury of health, is the modeling dosage that is fit to; Flash of light ERG can be used as the quantizating index of retina injury degree this model is assessed.
The method that 2 the present invention of test example screen the medicine of treatment retinal degeneration
Concrete steps:
A, adopt sodium iodate to be applied to Rhesus Macacus, set up Rhesus Macacus retinal degeneration model by the method for embodiment 1;
B, drug candidate is applied to the degeneration model;
C, observe the influence situation of drug candidate, and mark, estimate the medicine of potential treatment retinal degeneration the quantizating index of retinal degeneration, degree of injury.
Claims (6)
1. Rhesus Macacus retinal degeneration Preparation of model method is characterized in that: with dosage is that the sodium iodate of 15-75mg/kg is applied to Rhesus Macacus.
2. Rhesus Macacus retinal degeneration Preparation of model method according to claim 1, it is characterized in that: the dosage of described sodium iodate is 50mg/kg.
3. the purposes of animal model in the medicine of screening treatment retinal degeneration of claim 1 or the preparation of 2 described methods.
4. method of medicine of screening the treatment retinal degeneration, it comprises the steps:
A, be applied to Rhesus Macacus, set up Rhesus Macacus retinal degeneration model with sodium iodate;
B, drug candidate is applied to the degeneration model;
C, observe the influence situation of drug candidate, and mark, estimate the medicine of potential treatment retinal degeneration the quantizating index of retinal degeneration, degree of injury.
5. the method for screening of medicaments according to claim 4, it is characterized in that: the application dosage of the sodium iodate described in the step a is: 15-75mg/kg.
6. the method for screening of medicaments according to claim 5, it is characterized in that: the application dosage of the sodium iodate described in the step a is: 50mg/kg.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103720714A (en) * | 2014-01-10 | 2014-04-16 | 南京医科大学第一附属医院 | Production method of model for retinitis pigmentosa disease of primate |
| CN105943548A (en) * | 2016-06-06 | 2016-09-21 | 同济大学 | Retinal degeneration model and drug screening method thereof |
| CN106822177A (en) * | 2017-03-14 | 2017-06-13 | 昭衍（苏州）新药研究中心有限公司 | A kind of preparation method of the retinal pigment degeneration model of rabbit |
| CN113057142A (en) * | 2021-03-30 | 2021-07-02 | 四川大学华西医院 | Method for constructing intraretinal and/or subretinal fibrosis animal model |
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-
2010
- 2010-03-24 CN CN 201010131148 patent/CN102198278B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| 宫媛媛等: "碘酸钠诱导大鼠视网膜变性的病理学特征", 《中国眼底病论坛·全国眼底病专题学术研讨会论文汇编》 * |
| 王凯等: "碘酸钠诱导兔视网膜和视神经跨神经元变性的研究", 《眼科研究》 * |
| 胡小凤等: "碘酸钠静脉注入后兔眼图形VEP的改变", 《眼科研究》 * |
Cited By (8)
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| CN103720714B (en) * | 2014-01-10 | 2016-08-24 | 南京医科大学第一附属医院 | A kind of method making primate retina pigment denaturation model |
| CN105943548A (en) * | 2016-06-06 | 2016-09-21 | 同济大学 | Retinal degeneration model and drug screening method thereof |
| CN105943548B (en) * | 2016-06-06 | 2019-01-25 | 同济大学 | A kind of retinal degeneration model and its drug screening method |
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