CN103830721A - Derivant of establishing rheumatoid arthritis animal model, preparation and application thereof - Google Patents
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Abstract
The invention discloses a derivant of establishing a rheumatoid arthritis animal model, and preparation and application thereof; the derivant comprises II type collagen, aluminium hydroxide, glacial acetic acid and complete freund adjuvant; the derivant can successfully form the rheumatoid arthritis animal model, can substantially boost immunization reaction, can keep arthritis symptom to last for a longer time, so clinic symptom and pathology change of the animal model can be more close to real rheumatoid arthritis; the preparation method of the derivant is simple in operation, and convenient for promotion and application; the derivant is used for establishing the rheumatoid arthritis animal model, is simple and safe in realization, good in stable reliability, high in molding success rate, and has excellent application prospect.
Description
Technical field
The invention belongs to biological field, relate to a kind of derivant and preparation and application thereof of setting up rheumatoid arthritis animal model.
Technical background
Rheumatoid arthritis (rheumatoid arthritis, RA) be the chronic autoimmune disease taking symmetry polyarthritis as main clinical manifestation, the prevalence of China is 0.32%-0.36%, is to cause labour force to lose one of principal disease with disabled.
RA is a kind of multiple joints of whole body of invading, pathological characteristics is intraarticular synovitis, early stage is synovial membrane hyperemia, synovial membrane is loose, the newborn fiber of chronic inflammatory disease vascular tissue rolls up, and causes synovial membrane irregular thickening, and fine hair plumpness is charged into formation pannus in articular cavity, and then erosion damage ossa articularia cortex and cartilaginous tissue, cause eventually ankylosis, deformity and afunction; That the course of disease is is chronic, carrying out property, aggressive, treatment not in time, and disease progression is rapid, and joint deformity is a kind of infringement of irreversibility.Its pathogenesis is very complicated, illustrates not yet completely so far.
RA is carried out to a large amount of animal experiment studies both at home and abroad, and has formed some comparatively ripe animal models:
(1) adjuvant-induced arthritis (AA) model, this is owned by France in gross stress reaction, lacks chronic pathology process, and pathological changes has certain self limiting.
(2) collagen (CIA) arthritis model, has typical arthritis sign, and sickness rate is than using separately Freund's complete adjuvant injection to significantly improve, and the similar RA such as CIA model clinical symptoms, pathological change.
(3) arthritis model of egg protein induction, though this model success rate is high, has different in amynologic index and course of disease feature and RA.
(4) disease combination model, this model is not perfect, disease attribute evaluation difficulty, the problems such as stability, poor reliability, and do not adopted by clinical basic.
To sum up, CIA model is the ideal model of research and screening treatment RA medicine, and Ye Shi U.S. food Drug Administration is used for the treatment of the animal model of RA drug screening.
II Collagen Type VI (CII) is current comparatively generally acknowledged articular cartilage specific antigen, and the monokine il-1 (IL-1) relevant to cartilage destruction, the diacrisis of tumor necrosis factor α (TNF-α) are the key links of osteoarthrosis morbidity.
Due to the restriction of experiment condition, the zoopery object of in the past studying CIA model is generally limited to rat, mice etc., and primate is relatively less, but the animals such as rat are huge with mankind's difference after all.Small animal model can not be fine pathogenic process and the lesion degree of simulation people RA, increasing the weight of and alleviating of the state of an illness can not be described.In addition, existing building the derivant using in CIA model because immunoreation degree is limited, and inflammatory reaction hold time shorter, clinical symptoms and the pathological change that still can not well simulate rheumatoid arthritis.
Summary of the invention
For the problems referred to above, the invention provides a kind of brand-new derivant of setting up rheumatoid arthritis animal model.
The present invention is achieved through the following technical solutions:
Set up a derivant for rheumatoid arthritis animal model, described derivant is mixed emulsion, comprises II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant.Wherein II Collagen Type VI, as articular cartilage specific antigen, can be induced rheumatoid arthritis relevant symptoms; Glacial acetic acid is convenient to each component as solvent and is formed uniform mixed emulsion; Aluminium hydroxide and complete Freund's adjuvant are mutually collaborative, significantly booster immunization reaction, and aluminium hydroxide can also make arthritic symptom maintain the longer time simultaneously, makes the clinical symptoms of animal model and pathological change more approach real rheumatoid arthritis.
As optional mode, in above-mentioned derivant, described II Collagen Type VI is cattle II Collagen Type VI.Cattle II Collagen Type VI easily obtains, and adopts the similarity degree of cattle II Collagen Type VI symptom higher in the time building animal model using Rhesus Macacus as animal pattern.
As optional mode, in above-mentioned derivant, described II Collagen Type VI final concentration is 2-4 mg/ml, and aluminium hydroxide final concentration is 0.5-1.5 mg/ml.Adopt this concentration range both can induce and produce enough significantly symptom, avoid again introducing a large amount of derivants in animal pattern body, reduce the misery of animal while introducing derivant, improve animal welfare.
As optional mode, in above-mentioned derivant, described II Collagen Type VI final concentration is 3 mg/ml, and aluminium hydroxide final concentration is 1mg/ml.
The present invention also provides a kind of method of preparing above-mentioned derivant, comprises the following steps:
1) preparation of glacial acetic acid solution: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) preparation of collagen solution: II Collagen Type VI is joined in the glacial acetic acid solution that step 1) makes, after piping and druming evenly, preserve more than 8 hours in 4 DEG C of refrigerators, make II Collagen Type VI solution;
3) preparation of collagen Emulsion: using step 2) in the II Collagen Type VI solution that makes drop to equal volume complete Freund's adjuvant (as optional mode, the temperature of described complete Freund's adjuvant is controlled at 4-8 DEG C) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, and 4 DEG C of Refrigerator stores are for subsequent use.
As optional mode, in the preparation method of above-mentioned derivant, described step 2) in the concentration of II Collagen Type VI solution be 4-8 mg/ml.
The present invention also provides a kind of method of setting up rheumatoid arthritis animal model, comprises the following steps:
1) back Intradermal sensitization: choose healthy animal pattern, along its skin of back subcutaneous injection derivant of the present invention, in every animal pattern, the injected dose of II Collagen Type VI is 1-2mg;
2) raise and observe 40 days;
3) animal model of the acquisition rheumatoid arthritis positive.
As optional mode, in the above-mentioned method of setting up rheumatoid arthritis animal model, in described step 1), the injected dose of II Collagen Type VI is 1.5mg.
As optional mode, in the above-mentioned method of setting up rheumatoid arthritis animal model, in described step 1), the point of point multiple diverse locations is injected, and the injection volume of each point is all below 0.2mL.Make on the one hand derivant Multipoint Uniform distribute, be conducive to symptom and produce, on the other hand, reduce the injection volume of a single point, reduce animal suffering, improve animal welfare.
As optional mode, in the above-mentioned method of setting up rheumatoid arthritis animal model, choose non-human primates Rhesus Macacus as animal pattern, 2-5 year, weight range when use: 2.5-4.0kg.Rhesus Macacus and the mankind belong to primates together, and its biological characteristics and the mankind are close, according to modelling verification test of the present invention; the clinical manifestation of Rhesus Macacus CIA model; pathology etc. are similar to people RA, show chronic arthritis disease and osteoclasia, can evaluate the regulating and controlling effect of antiinflammatory, bone protection aspect simultaneously.The rheumatoid arthritis animal model that adopts method of the present invention to set up, experimental monkey groups shows the major lesions feature of collagen induced arthritis (CIA) model: D12-D15 after injection derivant, there is inflammatory reaction, injection site, back skin ulceration is hemorrhage, clinical manifestation is that grazing speed slows down, weight loss, biped dystropy D15-D25, and disease is rapid, be slow in action and do not like and climb articulations digitorum manus redness then, serological index is abnormal, and (CPR, NEUT significantly raise; ALB significantly reduces); D20-D40, joint serious swelling, is slow in action stiff, can not independently search for food, and needs the symptoms such as artificial feed to occur.The x-ray sign of Rhesus Macacus collagen induced arthritis (CIA) model that employing this method builds is similar to clinical RA, both hands and biped metacarpophalangeal joints (MCP)/metatarsophalangeal joints and nearly section articulations digitorum manus (PIP)/nearly section toe joint occur the initial stage or mid-term characteristics of lesion, the performance narrow and bone erosion of periarticular soft tissues swelling, regional osteoporosis, joint space of getting involved has specificity most with edge, joint bone erosion; pathological changes is taking hands or sufficient far-end, proximal interphalangeal joint as main, and wherein joints of foot destroys prior to joints of hand.Continue subsequently to observe to D40, joint serious swelling, model of rheumatoid arthritis is successfully set up.
Disclosed all features in this description, or step in disclosed all methods or process, except mutually exclusive feature and/or step, all can combine by any way.
Beneficial effect of the present invention:
1. derivant of the present invention can successfully build rheumatoid arthritis animal model, and significantly booster immunization reaction, can also make arthritic symptom maintain the longer time, make the clinical symptoms of animal model and pathological change more approach real rheumatoid arthritis.
2. described in, the preparation method of derivant is simple to operate, easy to utilize.
3. described in, set up the method for rheumatoid arthritis animal model; simple; safety; reliability is good, and modeling success rate is high, has a good application prospect; little to animal injury; and symptom and real rheumatoid arthritis similarity degree are high, show chronic arthritis disease and osteoclasia, and gained model can be evaluated the regulating and controlling effect of antiinflammatory, bone protection aspect simultaneously.
4. by setting up rheumatoid arthritis primate model; can be used for studying the pathogenesis of RA, screening bone protection medicine, explores principle of drug action; even evaluate regulating and controlling effect and the mechanism of action of related drugs to this kind of disease, tool is of great significance.
brief description of the drawings:
Fig. 1 is hematology and the Analysis of Biochemical result that in embodiment 5, the corresponding animal of experimental group 3 is relevant to inflammation.
Fig. 2 is the corresponding animal finger-joint immunity of experimental group 3 before and after look result in embodiment 5, and A, B are finger shape appearance figure before and after immunity; C, D point X ray picture before and after being respectively immunity.
Fig. 3 is the corresponding animal foot of experimental group 3 joint immunity before and after look result in embodiment 5, and A, B are foot shape appearance figure before and after immunity; C, D are respectively immunity front and back foot X ray picture.
Fig. 4 is the corresponding joint of animal pathological changes of experimental group 3 slice map in embodiment 5, and wherein (amplification is 10 for A, HE dyeing) characterizes synovial cell's fine hair shape hypertrophy, companion's cell infiltration (←); (amplification is 10 for B, HE dyeing) characterizes articular cartilage surface damage and local fibrosis (→).(amplification is 5 for C, HE dyeing) characterizes subcutaneous inflammatory disorders: cartilage surface and osseous tissue erosion.(amplification is 20 for D, HE dyeing) characterizes cartilage inflammatory disorders: cartilage surface is destroyed, by fibroblast is substituted.。
Fig. 5 is the preparation flow figure of derivant of the present invention.
detailed description of the invention:
Detailed description of the invention is by the following examples described in further detail foregoing of the present invention again.But this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following example.Do not departing from any amendment of making within the spirit and principles in the present invention, and being equal to of making according to ordinary skill knowledge and customary means replace or improve, all should be included in protection scope of the present invention.Raw materials usedly in following examples all can buy from the market.
embodiment 1prepare derivant (preparation flow as shown in Figure 5)
1) preparation glacial acetic acid solution: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) preparation collagen solution: II Collagen Type VI is joined in the glacial acetic acid solution that step 1) makes, after piping and druming evenly, preserve more than 8 hours in 4 DEG C of refrigerators, make II Collagen Type VI solution, described II Collagen Type VI can be the conventional II Collagen Type VIs such as cattle, pig, people;
3) preparation collagen Emulsion: using step 2) in the II Collagen Type VI solution that makes drop to equal volume complete Freund's adjuvant (as optional mode, the temperature of described complete Freund's adjuvant is controlled at 4-8 DEG C) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, the mixed emulsion that acquisition contains II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, 4 DEG C of Refrigerator stores are for subsequent use.
embodiment 2prepare derivant
1) preparation glacial acetic acid solution: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) preparation collagen solution: II Collagen Type VI is joined in the glacial acetic acid solution that step 1) makes, and the II Collagen Type VI solution that the concentration that is mixed with collagen solution is 4mg/ml, after piping and druming evenly, preserves 8 hours in 4 DEG C of refrigerators;
3) preparation collagen Emulsion: using step 2) in the II Collagen Type VI solution that makes drop to equal volume complete Freund's adjuvant (as optional mode, the temperature of described complete Freund's adjuvant is controlled at 4 DEG C of left and right) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, the mixed emulsion that acquisition contains II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, wherein II Collagen Type VI final concentration is 2 mg/ml, and aluminium hydroxide final concentration is 0.5 mg/ml, and 4 DEG C of Refrigerator stores are for subsequent use.
embodiment 3prepare derivant
1) preparation glacial acetic acid solution: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) preparation collagen solution: II Collagen Type VI is joined in the glacial acetic acid solution that step 1) makes, and the II Collagen Type VI solution that the concentration that is mixed with collagen solution is 8mg/ml, after piping and druming evenly, preserves 12 hours in 4 DEG C of refrigerators;
3) preparation collagen Emulsion: using step 2) in the II Collagen Type VI solution that makes drop to equal volume complete Freund's adjuvant (as optional mode, the temperature of described complete Freund's adjuvant is controlled at 4 DEG C of left and right) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, the mixed emulsion that acquisition contains II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, wherein II Collagen Type VI final concentration is 4 mg/ml, and aluminium hydroxide final concentration is 1.5 mg/ml, and 4 DEG C of Refrigerator stores are for subsequent use.
embodiment 4prepare derivant
1) preparation glacial acetic acid solution: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) preparation collagen solution: cattle II Collagen Type VI is joined in the glacial acetic acid solution that step 1) makes, and the II Collagen Type VI solution that the concentration that is mixed with collagen solution is 6mg/ml, after piping and druming evenly, preserves 12 hours in 4 DEG C of refrigerators;
3) preparation collagen Emulsion: using step 2) in the II Collagen Type VI solution that makes drop to equal volume complete Freund's adjuvant (as optional mode, the temperature of described complete Freund's adjuvant is controlled at 4 DEG C of left and right) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, the mixed emulsion that acquisition contains II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, wherein II Collagen Type VI final concentration is 3 mg/ml, and aluminium hydroxide final concentration is 1 mg/ml, and 4 DEG C of Refrigerator stores are for subsequent use.
embodiment 5the foundation of rheumatoid arthritis animal model
Choosing of animal
Choose non-human primates Rhesus Macacus as animal pattern, 3-5 year, weight range when use: 2.5-4.0kg.Adopt necklace label and the individual animals numbering of listing simultaneously.
(1) the selected front following items inspection of animal: two bacillus subtuberculosises tests, the inspection of feces parasite and health check-up etc., reject the animal that is not suitable for testing.Audit report comprises clinical examination and lab testing content, all carries out with reference to national standard.
(2) the quarantine domestication of animal after selected, 1 week, thes contents are as follows totally:
I. animal appearance physical examination, content mainly comprises: observe the appearance, the bodily form, action, body temperature, breathing of animal, by hair, nose, mouth, eye, ear, genitals, urine, feces etc.
Ii. the parasite to animal, antibacterial and microorganism detect: comprise pathogen, virus checking in animal body, endoparasite and ectoparasite inspection.
Iii. quarantine between domestication every day at the upper and lower noon twice totally pair of quarantine animal observe, content mainly comprises: animal appearance, feces, the situation etc. of ingesting.
The foundation of model
Choose satisfactory healthy Rhesus Macacus as animal pattern, test grouping by table 1,5 every group, inject derivant of the present invention along subcutaneous point of 10 diverse locations of its skin of back, the injection volume of each position is all controlled at below 0.2mL;
Table 1 experiment grouping information slip
| Group | Experimental group 1 | Experimental group 2 | Experimental group 3 | Experimental group 4 | Experimental group 5 | Contrast groups 1 | Contrast groups 2 |
| Collagen dosage (mg) | 1.5 | 1.5 | 1.5 | 1 | 2 | 1.5 | 1.5 |
| Derivant | Prepared by embodiment 2 | Prepared by embodiment 3 | Prepared by embodiment 4 | Prepared by embodiment 4 | Prepared by embodiment 4 | CII+ complete Freund's adjuvant | CII |
Breeding observing
Raise and observe 40 days at non-barrier system environment, the conventional nutrient component determining of feedstuff: supply of forage business provide, and determines within specified standard scope (with reference to National Standard of the People's Republic of China GB14924.8-2001).
Diagnostic mode evaluation methodology and result
Comprise serum biochemistry mark and hematological examination, x-ray diagnostic imaging, pathological examination etc.Wherein every test result the best of the 3rd experimental group, symptom approaches RA most, and it is the longest to hold time.Below only describe with the observed result of the 3rd experimental group.
1, serum biochemistry mark and the hematological examination factor mainly comprise neutrophil percentage (NEUT), c reactive protein (CRP), albumin (ALB) etc.
Hematological examination and serum biochemistry mark review time are quanrantine 1 time, after injectable collagen 1 time weekly.Hematological examination is used instrument to be, Sysmex XT-2000i type five blood cell analyzer of classifying.Serum biochemistry mark uses instrument to be: Beckman Kuerle automatic clinical chemistry analyzer model:SYNCHRON CX4 PRO.
Test result shows:
Organize collagen-induced before (D0) CRP, ALB, NEUT be Rhesus Macacus range of normal value; Before (see figure 1) is collagen-induced, (D0) CRP is that when D7, CRP significantly raises (69mg/L) in normal level (< 1mg/L), when D 21-D40, and CPR sharply raise (> 160 mg/L); Simultaneously visible injectable collagen D7-D40, ALB continuous decrease, neutrophil percentage significantly raises; Injectable collagen D35, ALB is only that 12.2 g/L, neutrophil percentage rise to 79.9%.
2, X ray image diagnosis
Within after injectable collagen 23-25 days, irradiate X-ray; .Use equipment for Philips all direction multifunctional veneer digital imaging system, model be Digital Diagnost VM.
Diagnostic imaging main points: 1 soft tissue of joint swelling; 2 joint spaces narrow, articular surface marginal bone corrodes; Under 3 bone articular surfaces, sclerotin folliculus shape destroys; 4 osteoporosises; 5 joint deformities and ankylosis.Bone erosion starts from the edge of articular cartilage, and marginality corrodes, and is the early stage important sign of RA.
The basic x-ray sign of RA: the narrow and bone erosion of periarticular soft tissues swelling, regional osteoporosis, joint space, has specificity most with edge, joint bone erosion.
Hands 1-5 proximal interphalangeal joint hyperosteogeny, sclerosis, articular surface bone destruction, joint space shows fuzzy, obvious stenosis, surrounding soft tissue's shade mild swelling thickens, and joint space has no obvious stenosis (see figure 2).The nearly toe joint hyperosteogeny of foot 2-5, sclerosis, articular surface bone destruction, joint space obvious stenosis, surrounding soft tissue's mild swelling, 2-5 toe joint far away joint space shows fuzzy, the suspicious destruction of articular surface sclerotin, joint space has no obvious stenosis (see figure 3).
3, liver histopathological analysis
According to X ray and sundry item check result, collagen-induced D40, in euthanasia, gets animal pathological changes extremities joint and completes pathological examination.
Tissue is fixed through 4% paraformaldehyde phosphate buffer, after then using the decalcification of formic acid hydrochloric acid decalcifying Fluid completely, pathologic sampling, dehydration, paraffin embedding, cuts 5um thin slice, HE dyeing, and microscopic examination is also taken pictures.
Arthritis pathology pathological changes: visible joint, joint shallow-layer hyaline cartilage is destroyed, for fibrous connective tissue substitutes; The a large amount of cell infiltration of joint capsule undertissue.
Synovitis pathological changes: (Fig. 4-A and 4-B, HE × 10) synovial cell's fine hair shape hypertrophy, companion's cell infiltration (
).Articular cartilage surface damage, local fibrosis (
).
Subcutaneous inflammatory disorders: (Fig. 4-C, HE × 5) cartilage surface and osseous tissue erosion.
Cartilage inflammatory disorders: (Fig. 4-D, HE × 20) cartilage surface is destroyed, by fibroblast is substituted.
In sum, rheumatoid arthritis animal model modeling success prepared by the present invention, good stability, the evaluation of the biotechnology new drug that can cannot complete for rodent models.
In the present embodiment, the modeling success rate of experimental group 1-5 is 100%, and each experimental group group interpolation heteropole is little, has difference in various degree between group in duration of symptoms, the duration of symptoms of experimental group 1,2,4,5 is about 1 month, and the duration of symptoms of experimental group 3 can reach 2 months.But clinical symptoms and the phenotype that all can successfully simulate rheumatoid arthritis of each experimental group.Wherein with resultant effect the best of experimental group 3.Contrast groups 1,2 symptom similarity degree compared with each experimental group is obviously poor, and a little less than immunoreation, duration of symptoms is short, and modeling success rate is lower is about below 30%.
The foregoing is only the preferred embodiments of the present invention, is only illustrative for the purpose of the present invention, and nonrestrictive; Those of ordinary skill in the art understand, and in the spirit and scope that limit, can carry out many changes to it in the claims in the present invention, amendment, and even equivalence is changed, but all will fall into protection scope of the present invention.
Claims (9)
1. a derivant of setting up rheumatoid arthritis animal model, is characterized in that, comprises II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant.
2. the derivant of setting up rheumatoid arthritis animal model according to claim 1, is characterized in that, described II Collagen Type VI is cattle II Collagen Type VI.
3. the derivant of setting up rheumatoid arthritis animal model according to claim 1, is characterized in that, described II Collagen Type VI final concentration is 2-4 mg/ml, and aluminium hydroxide final concentration is 0.5-1.5 mg/ml.
4. the derivant of setting up rheumatoid arthritis animal model according to claim 1, is characterized in that, described II Collagen Type VI final concentration is 3 mg/ml, and aluminium hydroxide final concentration is 1mg/ml.
5. according to the preparation method of the derivant described in any one in claim 1-4, it is characterized in that, comprise the following steps:
1) preparation of glacial acetic acid solution: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) preparation of collagen solution: II Collagen Type VI is joined in the glacial acetic acid solution that step 1) makes, after piping and druming evenly, preserve more than 8 hours in 4 DEG C of refrigerators, make II Collagen Type VI solution;
3) preparation of collagen Emulsion: by step 2) in the II Collagen Type VI solution that makes drop in equal volume complete Freund's adjuvant, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, 4 DEG C of Refrigerator stores are for subsequent use.
6. a method of setting up rheumatoid arthritis animal model, is characterized in that, comprises the following steps:
1) back Intradermal sensitization: choose healthy animal pattern, as the derivant as described in any one in claim 1-4, in every animal pattern, II Collagen Type VI injected dose is 1-2mg along its skin of back subcutaneous injection;
2) raise and observe 40 days;
3) animal model of the acquisition rheumatoid arthritis positive.
7. the method for setting up rheumatoid arthritis animal model according to claim 6, is characterized in that, in described step 1), the injected dose of II Collagen Type VI is 1.5mg.
8. the method for setting up rheumatoid arthritis animal model according to claim 6, is characterized in that: in described step 1), the point of point multiple diverse locations is injected, and the injection volume of each point is all below 0.2mL.
9. the method for setting up rheumatoid arthritis animal model according to claim 6, is characterized in that: choose non-human primates Rhesus Macacus as animal pattern, 2-5 year, weight range when use: 2.5-4.0kg.
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| CN110476890A (en) * | 2019-08-07 | 2019-11-22 | 南通大学 | A kind of preparation method, application and the evaluation method of Collagen-induced Arthritis animal model |
| CN114916501A (en) * | 2022-05-16 | 2022-08-19 | 四川大学华西医院 | Application of diphenylene iodonium chloride in model molding of rheumatoid arthritis and model molding method of rheumatoid arthritis |
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| CN106177999A (en) * | 2016-08-04 | 2016-12-07 | 南京大学 | The method for building up of a kind of joint of vertebral column inflammation mouse model knocking out technology based on conditionality and purposes |
| CN106177999B (en) * | 2016-08-04 | 2019-04-16 | 南京大学 | A kind of method for building up and purposes of the joint of vertebral column inflammation mouse model knocking out technology based on conditionity |
| CN110476890A (en) * | 2019-08-07 | 2019-11-22 | 南通大学 | A kind of preparation method, application and the evaluation method of Collagen-induced Arthritis animal model |
| CN115281149A (en) * | 2021-05-12 | 2022-11-04 | 四川大学华西医院 | Preclinical rheumatoid arthritis (Pre-RA) mouse model |
| CN114916501A (en) * | 2022-05-16 | 2022-08-19 | 四川大学华西医院 | Application of diphenylene iodonium chloride in model molding of rheumatoid arthritis and model molding method of rheumatoid arthritis |
| CN114916501B (en) * | 2022-05-16 | 2023-01-31 | 四川大学华西医院 | Application of diphenylene iodonium chloride in model molding of rheumatoid arthritis and model molding method of rheumatoid arthritis |
| CN115413625A (en) * | 2022-08-22 | 2022-12-02 | 郑咏秋 | Rheumatoid coronary heart disease animal model and construction method and application thereof |
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