CN103830721B - A kind of derivant and preparation and application thereof of setting up rheumatoid arthritis animal model - Google Patents
A kind of derivant and preparation and application thereof of setting up rheumatoid arthritis animal model Download PDFInfo
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- CN103830721B CN103830721B CN201410057939.1A CN201410057939A CN103830721B CN 103830721 B CN103830721 B CN 103830721B CN 201410057939 A CN201410057939 A CN 201410057939A CN 103830721 B CN103830721 B CN 103830721B
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Abstract
The invention discloses a kind of derivant and preparation and application thereof of setting up rheumatoid arthritis animal model.Described derivant comprises II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, can successfully build rheumatoid arthritis animal model, and can significantly react by booster immunization, the time that arthritic symptom can also be made to remain longer, make the clinical symptoms of animal model and pathological change closer to real rheumatoid arthritis.The preparation method of described derivant is simple to operate, easy to utilize.Described derivant is used for setting up rheumatoid arthritis animal model, simple, safety, and reliability is good, and modeling success rate is high, has a good 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 (rheumatoidarthritis, RA) be the chronic auto-immune disease that is main clinical manifestation with symmetry polyarthritis, the prevalence of China is 0.32%-0.36%, is to cause disability and one of disabled principal disease.
RA is a kind of infringement multiple joint of whole body, pathological characteristics is intraarticular synovitis, early stage is that synovial membrane is congested, synovial membrane loosens, the newborn fibrovascular tissue of chronic inflammatory disease rolls up, and cause synovial membrane irregular thickening, fine hair plumpness is charged in articular cavity and formed pannus, and then erosion damage ossa articularia cortex and cartilaginous tissue, cause ankylosis, deformity and afunction eventually; The course of disease is chronic, Progressive symmetric erythrokeratodermia, aggressive, treats 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.
A large amount of animal experiment studies is carried out to RA both at home and abroad, and has defined some comparatively ripe animal models:
(1) adjuvant-induced arthritis (AA) model, this is owned by France in gross stress reaction, and lack chronic pathology process, pathological changes has certain self limiting.
(2) Collagen (CIA) arthritis model, has typical arthritis sign, and sickness rate ratio is used alone Freund's complete adjuvant injection and significantly improves, 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 problem such as stability, poor reliability, and is 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 the articular cartilage specific antigen of comparatively generally acknowledging at present, the diacrisis of the monokine il-1 (IL-1) relevant to cartilage destruction, tumor necrosis factor α (TNF-α), is the key link of osteoarthrosis morbidity.
Due to the restriction of experiment condition, the zoopery object 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 simulate pathogenic process and the lesion degree of people RA very well, can not describe increasing the weight of and alleviating of the state of an illness.In addition, the existing derivant used in structure CIA model is due to immunoreation limitation, and inflammatory reaction is held time shorter, still well can not simulate clinical symptoms and the pathological change of rheumatoid arthritis.
Summary of the invention
For the problems referred to above, the invention provides a kind of derivant setting up rheumatoid arthritis animal model completely newly.
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 is as articular cartilage specific antigen, can induced rheumatoid arthritis related 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 worked in coordination with mutually, and can significantly react by booster immunization, aluminium hydroxide can also make the time that arthritic symptom remains longer simultaneously, makes the clinical symptoms of animal model and pathological change closer to real rheumatoid arthritis.
Alternately, 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 when building animal model using Rhesus Macacus as animal pattern.
Alternately, in above-mentioned derivant, described II Collagen Type VI final concentration is 2-4mg/ml, and aluminium hydroxide final concentration is 0.5-1.5mg/ml.Adopt this concentration range both can induce generation enough significantly symptom, avoid again introducing a large amount of derivants in animal pattern body, reduce the misery of animal when introducing derivant, improve animal welfare.
Alternately, in above-mentioned derivant, described II Collagen Type VI final concentration is 3mg/ml, and aluminium hydroxide final concentration is 1mg/ml.
Present invention also offers a kind of method preparing above-mentioned derivant, comprise the following steps:
1) preparation of glacial acetic acid solution: get glacial acetic acid, add distilled water, is 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 obtained glacial acetic acid solution of step 1), after piping and druming evenly, preserve more than 8 hours in 4 DEG C of refrigerators, obtained II Collagen Type VI solution;
3) preparation of collagen emulsion: by step 2) in II obtained Collagen Type VI solution drop to equal volume complete Freund's adjuvant (alternately, the temperature of described complete Freund's adjuvant controls 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.
Alternately, in the preparation method of above-mentioned derivant, described step 2) in the concentration of II Collagen Type VI solution be 4-8mg/ml.
Present invention also offers a kind of method setting up rheumatoid arthritis animal model, comprise 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 rheumatoid arthritis positive is obtained.
Alternately, set up in the method for rheumatoid arthritis animal model above-mentioned, in described step 1), the injected dose of II Collagen Type VI is 1.5mg.
Alternately, set up in the method for rheumatoid arthritis animal model above-mentioned, in described step 1), the point of point multiple diverse location is injected, and the injection volume of each point is all at below 0.2mL.On the one hand derivant Multipoint Uniform is distributed, be conducive to symptom and produce, on the other hand, reduce the injection volume of a single point, reduce animal suffering, improve animal welfare.
Alternately, set up in the method for rheumatoid arthritis animal model above-mentioned, choose non-human primates Rhesus Macacus as animal pattern, 2-5 year, weight range during use: 2.5-4.0kg.Rhesus Macacus and the mankind belong to primates together, and its biological characteristics is close with the mankind, and modelling verification according to the present invention is tested; the clinical manifestation of Rhesus Macacus CIA model; pathology etc. are similar to people RA, show chronic joint inflammation and osteoclasia, can evaluate the regulating and controlling effect of antiinflammatory, bone protection aspect simultaneously.Adopt the rheumatoid arthritis animal model of method establishment of the present invention, experimental monkey groups shows the major lesions feature of collagen induced arthritis (CIA) model: D12-D15 after injection derivant, there is inflammatory reaction, dorsal injection area skin festers hemorrhage, clinical manifestation is that grazing speed slows down, and body weight reduces, 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 adopting this method to build 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, narrow and the bone erosion in performance affected joints surrounding soft tissue swelling, regional osteoporosis, joint space has specificity most with joint margins bone erosion; pathological changes is based on hands or sufficient far-end, proximal interphalangeal joint, 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.
All features disclosed in this description, or the 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 can significantly react by booster immunization, the time that arthritic symptom can also be made to remain longer, make the clinical symptoms of animal model and pathological change closer to 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 high, show chronic joint inflammation and osteoclasia, gained model can evaluate 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 related drugs to the regulating and controlling effect of this kind of disease and mechanism of action, tool is of great significance.
accompanying drawing illustrates:
Fig. 1 is the hematology and Analysis of Biochemical result that in embodiment 5, the corresponding animal of experimental group 3 is relevant to inflammation.
Fig. 2 is experimental group 3 corresponding animal finger-joint immunity before and after look result in embodiment 5, and A, B are finger shape appearance figure before and after immunity; C, D are respectively finger X ray picture before and after immunity.
Fig. 3 is experimental group 3 corresponding animal foot 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 foot X ray picture before and after immunity.
Fig. 4 is experimental group 3 corresponding joint of animal pathological changes slice map in embodiment 5, and wherein (A, HE dye, and amplification is 10) characterizes synovial cell's villous hyperplasia, companion's cell infiltration (←); (B, HE dye, and amplification is 10) characterizes articular cartilage surface damage and local fibrosis (→).(C, HE dye, and amplification is 5) characterizes subcutaneous inflammatory disorders: cartilage surface and osseous tissue erosion.(D, HE dye, and amplification is 20) 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 that the scope of the above-mentioned theme of the present invention is only limitted to following example.Not departing from any amendment made within the spirit and principles in the present invention, and the equivalent replacement made according to ordinary skill knowledge and customary means or improvement, all should be included in protection scope of the present invention.Raw materials usedly in following examples all can to buy from the market.
embodiment 1prepare derivant (preparation flow as shown in Figure 5)
1) glacial acetic acid solution is prepared: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) prepare collagen solution: joined by II Collagen Type VI in the obtained glacial acetic acid solution of step 1), after piping and druming evenly, preserve more than 8 hours in 4 DEG C of refrigerators, obtained II Collagen Type VI solution, described II Collagen Type VI can be the conventional II Collagen Type VI such as cattle, pig, people;
3) prepare collagen emulsion: by step 2) in II obtained Collagen Type VI solution drop to equal volume complete Freund's adjuvant (alternately, the temperature of described complete Freund's adjuvant controls at 4-8 DEG C) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, obtain the mixed emulsion containing 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) glacial acetic acid solution is prepared: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) prepare collagen solution: joined by II Collagen Type VI in the obtained glacial acetic acid solution of step 1), the concentration being mixed with collagen solution is the II Collagen Type VI solution of 4mg/ml, after piping and druming evenly, preserve 8 hours in 4 DEG C of refrigerators;
3) prepare collagen emulsion: by step 2) in II obtained Collagen Type VI solution drop to equal volume complete Freund's adjuvant (alternately, the temperature of described complete Freund's adjuvant controls at about 4 DEG C) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, obtain the mixed emulsion containing II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, wherein II Collagen Type VI final concentration is 2mg/ml, and aluminium hydroxide final concentration is 0.5mg/ml, and 4 DEG C of Refrigerator stores are for subsequent use.
embodiment 3prepare derivant
1) glacial acetic acid solution is prepared: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) prepare collagen solution: joined by II Collagen Type VI in the obtained glacial acetic acid solution of step 1), the concentration being mixed with collagen solution is the II Collagen Type VI solution of 8mg/ml, after piping and druming evenly, preserve 12 hours in 4 DEG C of refrigerators;
3) prepare collagen emulsion: by step 2) in II obtained Collagen Type VI solution drop to equal volume complete Freund's adjuvant (alternately, the temperature of described complete Freund's adjuvant controls at about 4 DEG C) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, obtain the mixed emulsion containing II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, wherein II Collagen Type VI final concentration is 4mg/ml, and aluminium hydroxide final concentration is 1.5mg/ml, and 4 DEG C of Refrigerator stores are for subsequent use.
embodiment 4prepare derivant
1) glacial acetic acid solution is prepared: get glacial acetic acid, add distilled water, be diluted to the glacial acetic acid solution that concentration is 0.1mol/L;
2) prepare collagen solution: joined by cattle II Collagen Type VI in the obtained glacial acetic acid solution of step 1), the concentration being mixed with collagen solution is the II Collagen Type VI solution of 6mg/ml, after piping and druming evenly, preserve 12 hours in 4 DEG C of refrigerators;
3) prepare collagen emulsion: by step 2) in II obtained Collagen Type VI solution drop to equal volume complete Freund's adjuvant (alternately, the temperature of described complete Freund's adjuvant controls at about 4 DEG C) in, add aluminium hydroxide simultaneously, rapid stirring makes it fully emulsified, obtain the mixed emulsion containing II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant, wherein II Collagen Type VI final concentration is 3mg/ml, and aluminium hydroxide final concentration is 1mg/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 during use: 2.5-4.0kg.Simultaneously adopt necklace list label and individual animals numbering.
(1) the selected front following items inspection of animal: two bacillus subtuberculosises tests, the inspection of feces parasite and health check-up etc., rejects the animal being not suitable for carrying out testing.Audit report comprises clinical examination and lab testing content, all performs with reference to national standard.
(2) animal be selected in after quarantine domestication, totally 1 week, content was as follows:
I. animal appearance physical examination, content mainly comprises: observe the appearance of animal, the bodily form, action, body temperature, breathing, by hair, nose, mouth, eye, ear, genitals, urine, feces etc.
Ii. the parasite of animal, antibacterial and microorganism are detected: comprise pathogen in animal body, virus checking, endoparasite and ectoparasite inspection.
Iii. between quarantine domestication, every day at the upper and lower noon observes quarantine animal for twice totally, and content mainly comprises: animal appearance, feces, situation etc. of ingesting.
The foundation of model
Choose satisfactory healthy Rhesus Macacus as animal pattern, carry out experiment grouping by table 1, often organize 5, inject derivant of the present invention along its skin of back subcutaneous point of 10 diverse locations, the injection volume of each position all controls at below 0.2mL;
Grouping information slip tested by table 1
| 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 at non-barrier system environment and observe 40 days, feedstuff conventional nutrients measures: 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 biochemical markers thing and hematological examination, x-ray diagnostic imaging, pathological examination etc.Wherein every test result of the 3rd experimental group is best, and symptom is closest to RA, and it is the longest to hold time.Below be only described with the observed result of the 3rd experimental group.
1, serum biochemical markers thing and the hematological examination factor mainly comprise neutrophil percentage (NEUT), c reactive protein (CRP), albumin (ALB) etc.
Hematological examination and serum biochemical markers thing review time are quanrantine 1 time, after injectable collagen 1 time weekly.Hematological examination uses instrument to be that SysmexXT-2000i type five is classified blood cell analyzer.Serum biochemical markers thing use instrument is: Beckman Kuerle automatic clinical chemistry analyzer model:SYNCHRONCX4PRO.
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 for being in normal level (< 1mg/L), and during D7, CRP significantly raises (69mg/L), and during D21-D40, CPR sharply raises (> 160mg/L); Visible injectable collagen D7-D40, ALB continuous decrease simultaneously, neutrophil percentage significantly raises; Injectable collagen D35, ALB are only 12.2g/L, neutrophil percentage rises to 79.9%.
2, X ray image diagnosis
Within after injectable collagen 23-25 days, irradiate X-ray; .Use equipment is Philips all direction multifunctional veneer digital imaging system, and model is DigitalDiagnostVM.
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 namely marginality corrodes, and is the early stage important sign of RA.
RA basic x-ray sign: the narrow and bone erosion in periarticular soft tissues swelling, regional osteoporosis, joint space, has specificity most with joint margins bone erosion.
Hands 1-5 proximal interphalangeal joint hyperosteogeny, sclerosis, articular surface bone destruction, joint space display is fuzzy, and 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, the display of 2-5 toe joint far away joint space is fuzzy, and 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, is in euthanasia, gets animal pathological changes extremities joint and completes pathological examination.
Tissue is fixed through 4% paraformaldehyde phosphate buffer, and after then using the decalcification of formic acid hydrochloric acid decalcifying Fluid completely, pathologic sampling, dehydration, paraffin embedding, cut 5um thin slice, HE dyeing, microscopic examination is also taken pictures.
Arthritis pathology pathological changes: joint visible 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 villous hyperplasia, companion's cell infiltration (
).Articular cartilage surface is damaged, 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, may be used for the evaluation of the biotechnology new drug that rodent models cannot complete.
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 was at about 1 month, and the duration of symptoms of experimental group 3 can reach 2 months.But clinical symptoms and the phenotype that all successfully can simulate rheumatoid arthritis of each experimental group.Wherein best with the resultant effect of experimental group 3.Contrast groups 1,2 symptom similarity degree compared with each experimental group is obviously poor, and immunoreation is more weak, and duration of symptoms is short, and modeling success rate is lower is about less than 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 can carry out many changes in the spirit and scope that the claims in the present invention limit to it, amendment, and even equivalence is changed, but all will fall into protection scope of the present invention.
Claims (9)
1. set up a derivant for rheumatoid arthritis Rhesus Macacus animal model, it is characterized in that, comprise II Collagen Type VI, aluminium hydroxide, glacial acetic acid and complete Freund's adjuvant.
2. the derivant setting up rheumatoid arthritis Rhesus Macacus animal model according to claim 1, is characterized in that, described II Collagen Type VI is cattle II Collagen Type VI.
3. the derivant setting up rheumatoid arthritis Rhesus Macacus animal model according to claim 1, is characterized in that, described II Collagen Type VI final concentration is 2-4mg/ml, and aluminium hydroxide final concentration is 0.5-1.5mg/ml.
4. the derivant setting up rheumatoid arthritis Rhesus Macacus animal model according to claim 1, is characterized in that, described II Collagen Type VI final concentration is 3mg/ml, and aluminium hydroxide final concentration is 1mg/ml.
5. according to the preparation method of the derivant in claim 1-4 described in any one, it is characterized in that, comprise the following steps:
1) preparation of glacial acetic acid solution: get glacial acetic acid, add distilled water, is 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 obtained glacial acetic acid solution of step 1), after piping and druming evenly, preserve more than 8 hours in 4 DEG C of refrigerators, obtained II Collagen Type VI solution;
3) preparation of collagen emulsion: by step 2) in II obtained Collagen Type VI solution drop in equal volume complete Freund's adjuvant, add aluminium hydroxide, rapid stirring makes it fully emulsified simultaneously, and 4 DEG C of Refrigerator stores are for subsequent use.
6. set up a method for rheumatoid arthritis Rhesus Macacus animal model, it is characterized in that, comprise the following steps:
1) back Intradermal sensitization: choose healthy animal pattern, along its skin of back subcutaneous injection as the derivant in claim 1-4 as described in any one, in every animal pattern, II Collagen Type VI injected dose is 1-2mg;
2) raise and observe 40 days;
3) the Rhesus Macacus animal model of the rheumatoid arthritis positive is obtained.
7. the method setting up rheumatoid arthritis Rhesus Macacus 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 setting up rheumatoid arthritis Rhesus Macacus animal model according to claim 6, is characterized in that: in described step 1), the point of point multiple diverse location is injected, and the injection volume of each point is all at below 0.2mL.
9. the method setting up rheumatoid arthritis Rhesus Macacus animal model according to claim 6, is characterized in that: choose non-human primates Rhesus Macacus as animal pattern, 2-5 year, weight range during 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 |
| CN115281149A (en) * | 2021-05-12 | 2022-11-04 | 四川大学华西医院 | Preclinical rheumatoid arthritis (Pre-RA) mouse model |
| 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 |
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