CN105126075A - Establishment and application of rhesus monkey pulmonary fibrosis model - Google Patents
Establishment and application of rhesus monkey pulmonary fibrosis model Download PDFInfo
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- CN105126075A CN105126075A CN201510608667.4A CN201510608667A CN105126075A CN 105126075 A CN105126075 A CN 105126075A CN 201510608667 A CN201510608667 A CN 201510608667A CN 105126075 A CN105126075 A CN 105126075A
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Abstract
The invention discloses a method for preparing an animal pulmonary fibrosis model. According to the method, bleomycin is applied to a primate, the application dose of the bleomycin is 7 mg/Kg for each time, and application is performed once for each week and totally performed twice. The invention further provides a rhesus monkey pulmonary fibrosis model prepared with the method and an application of the rhesus monkey pulmonary fibrosis model. The rhesus monkey pulmonary fibrosis model is established successfully, an operation process is simple and easy to repeat, and the model can be used for new biotechnological drug evaluation which cannot be finished by a rodent model and has bright clinical application prospects.
Description
Technical field
The present invention relates to a kind of preparation method and its usage of Rhesus Macacus pulmonary fibrosis model.
Background technology
Pulmonary fibrosis is the major disease affecting human health, and pathogenic factor is failed to understand, so far not effective prevention and Therapeutic Method.Conventional mechanical ventilation, although hormone therapy can reduction of patient dyspnea to a certain extent, the situations such as hypoxemia, life-time service hormone therapy may bring out other diseases, and mechanical ventilation cost is higher, and operation not easily.Above Therapeutic Method all fundamentally can not cure pulmonary fibrosis, does not also improve survival rate.How to stop spreading of pulmonary fibrosis, effectively suppress fibroblastic growth in lung, fundamentally barrier fibers is the key point of research at present
The most frequently used the using a model of current pulmonary fibrosis research is Rodents.Because Rodents is far apart with the mankind in evolution, little to Clinical significance of MG with the experimental data that it obtains for object of study.The anatomical features of Rhesus Macacus, each system physiological function and similar to the mankind several with the reactivity of medicine to disease carry out treatment of fibrosis, and fibrocyte targeting removes the clinical front effectiveness of medicine and the Sensitivity animal of safety evaluatio.Utilize Rhesus Macacus pulmonary fibrosis model to evaluate Novel lung treatment of fibrosis medicine, research and development receptor target medicine effect of drugs are evaluated, and this has great importance to future therapeutic scheme and clinical practice.Because current pulmonary fibrosis medicine and method can not effectively control Fibrotic spreading, therefore setting up non-human primates pulmonary fibrosis model for newtype drug and targeted therapy provides evaluation to have urgent needs.
At present, there is no the report setting up Rhesus Macacus pulmonary fibrosis model.
Summary of the invention
Technical scheme of the present invention there is provided the method setting up Rhesus Macacus pulmonary fibrosis model.
The present invention provide firstly the purposes of bleomycin in the primate model for the preparation of screening treatment pulmonary fibrosis medicine, and wherein, the application dosage of bleomycin is: 7mg/Kg/ time, once in a week, uses twice altogether.
Preferably, the method for application of described bleomycin is atomization injection.
Invention additionally provides a kind of method preparing pulmonary fibrosis model animal model, and it is that bleomycin is applied to primate, and the application dosage of bleomycin is: 7mg/Kg/ time, once in a week, uses twice altogether.
Preferably, described primate is Rhesus Macacus.
Preferably, the method for application of described bleomycin is atomization injection.
Present invention also offers the pulmonary fibrosis model animal model that preceding method prepares.
Present invention also offers the application of aforementioned pulmonary fibrosis model animal model in screening pulmonary fibrosis medicine.
Present invention also offers a kind of method of screening the medicine for the treatment of pulmonary fibrosis, comprise the steps:
A, it is that bleomycin is applied to primate, and the application dosage of bleomycin is: 7mg/Kg/ time, once in a week, use twice altogether;
B, material standed for is applied to primate described in a step;
C, evaluate the medicine of potential treatment pulmonary fibrosis with the primate described in a step.
Preferably, primate described in a step is Rhesus Macacus.
Preferably, the method for application of described bleomycin described in a step is atomization injection
The inventive method successfully establishes Rhesus Macacus pulmonary fibrosis model, and operating process is simple, and easily repeat, may be used for the evaluation of the biotechnology new drug that rodent models cannot complete, potential applicability in clinical practice is good.
Below by way of detailed description of the invention, the present invention is described in further detail, but do not limit the present invention, those skilled in the art can make various change and distortion according to the present invention, only otherwise depart from spirit of the present invention, all should belong to the scope of claims of the present invention.
Accompanying drawing explanation
CT picture before Fig. 1 modeling
Fig. 2 modeling monkey CT picture
Detailed description of the invention
The preparation of embodiment 1 animal model of the present invention
1. materials and methods
1.1 experiment material
2 ?3 years old Rhesus Monkey 6, without B herpesvirus, monkey retrovirus, monkey leucovirus and immunodeficiency virus infection: purchased from Meishan animal Breeding base.
1.2 main agents
Bleomycin (BLM), purchased from Bai Dekang pharmacy, Beijing, Nippon Kayaku K. K produces.
Other medicines and consumptive material are purchased from love Deco skill.
1.3 main solution configurations
The configuration of bleomycin normal saline solution
Get 15mg to tire every bottle of bleomycin, inject 200 microlitre normal saline, form 75mg/ml bleomycin after dissolving and tire
1.4 key instrument
1 high-pressure atomization syringe: PennCentury
2 anesthetic machines: Excel210se
3CT machine: GEBRIVOCT325
4 anesthetic laryngopharyngoscopes
1.5 experimental technique
1.5.1 Rhesus Macacus lung CT detects, and before formal modeling, uses CT scan experimental monkey groups pulmonary, gets rid of the possibility of experimental monkey groups self pulmonary disease.
1). body weight when buying according to animal, inject ketamine (50mg/ml) by 15mg/kg
2). after Animal Anesthesia, weigh, record body weight
3). by fixing for Rhesus Macacus upper and lower extremities with on operating-table, lower limb posterior leg preserved skin exposes great saphenous vein, iodophor disinfection.
4). venoclysis needle puncture great saphenous vein, 5ml syringe blood drawing 5ml, extract out venoclysis needle, disinfecting cotton swab stop blooding, simultaneously by blood transfer in syringe to (yellow head coagulant pipe) in 5ml vacuum test tube, labelling, as later stage biochemistry detection.
5). the experimental monkey groups monkey under narcotism is transferred to CT machine scanning bed on, lie low, fixing limbs.
6). start CT machine scanning experimental monkey groups, scanning position is pulmonary, and record files.
7). after detection, the experimental monkey groups in anesthesia is transferred in rearging cage, observe until revive.
1.5.2 bleomycin air flue injection inducing lung fibrosis model, dosage regimen is 1 time weekly, uses 2 times altogether.
1). according to the body weight that last time measures, inject ketamine (50mg/ml) by 15mg/kg
2). after experimental monkey groups anesthesia, weigh, record body weight
3). with the experimental monkey groups body weight weighed for standard, bleomycin injection volume is 7mg/kg.If experimental monkey groups body weight is nkg, then need the bleomycin dosage injected to be 7nmg, the bleomycin normal saline solution of so taking is (7n/75ml).
4). use PennCentury syringe to draw the bleomycin normal saline solution configured.
5). lain low by Rhesus Macacus in operating-table, upper and lower extremities is fixed on operating-table, by padded for monkey shoulder, uses light source irradiation eyes, observes eyeball with or without motion, judges its narcotism.
6). hand-held PennCentury is atomized syringe, and injection head aims at fissure of glottis, inserts 2cm, and bleomycin normal saline solution in bolus syringe after inserting, until complete, extracts syringe out.
7). venoclysis needle puncture great saphenous vein, 5ml syringe blood drawing 5ml, extract out venoclysis needle, disinfecting cotton swab stop blooding, simultaneously by blood transfer in syringe to (yellow head coagulant pipe) in 5ml vacuum test tube, labelling, as later stage biochemistry detection.
8). observe monkey until will revive, transfer them to rearging cage.
9). observe animal activity ability twice weekly, and record.
1.5.3 after modeling, CT detects, and determines fibrosis lesion degree, and whether assessment models is successful.The same 1.5.1 of Examination on experimental operation.CT be detected on bleomycin injection after monthly.
2 experimental results
After modeling half a year, rhesus serum SP-D level obviously raises, and monkey mobility declines.CT detects, CT picture normal (Figure 1A, 1B) before 6 Rhesus Macacus modelings, after modeling, monkey lung CT all presents bilateral and fills the air distribution of net trellis shade, Honeycomb change (Fig. 2 A, 2B, 2C), be judged as pulmonary fibrosis pathological changes, pulmonary fibrosis model modeling success.
6 Rhesus Macacus set up pulmonary fibrosis model by the inventive method, and all success, illustrates that the inventive method can effectively set up Rhesus Macacus pulmonary fibrosis model, repeatable high.
To sum up, the inventive method successfully can set up river monkey pulmonary fibrosis model, and operating process is simple, easily repeats, may be used for the biotechnology new drug that rodent models cannot complete evaluation and.
Claims (10)
1. the purposes of bleomycin in the primate model for the preparation of screening treatment pulmonary fibrosis medicine, wherein, the application dosage of bleomycin is: 7mg/Kg/ time, once in a week, uses twice altogether.
2. purposes according to claim 1, is characterized in that: the method for application of described bleomycin is atomization injection.
3. prepare a method for pulmonary fibrosis model animal model, it is characterized in that: it is that bleomycin is applied to primate, the application dosage of bleomycin is: 7mg/Kg/ time, once in a week, use twice altogether.
4. preparation method according to claim 3, is characterized in that: described primate is Rhesus Macacus.
5. the preparation method according to claim 3 or 4, is characterized in that: the method for application of described bleomycin is atomization injection.
6. the pulmonary fibrosis model animal model that the method described in arbitrary one of claim 3-5 prepares.
7. the application of pulmonary fibrosis model animal model according to claim 6 in screening pulmonary fibrosis medicine.
8. screen a method for the medicine for the treatment of pulmonary fibrosis, comprise the steps:
A, it is that bleomycin is applied to primate, and the application dosage of bleomycin is: 7mg/Kg/ time, once in a week, uses twice altogether.
B, material standed for is applied to primate described in a step;
C, evaluate the medicine of potential treatment pulmonary fibrosis with the primate described in a step.
9. the method for screening of medicaments according to claim 8, is characterized in that: primate described in a step is Rhesus Macacus.
10. the method for screening of medicaments according to claim 8 or claim 9, is characterized in that: the method for application of described bleomycin described in a step is atomization injection.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107469069A (en) * | 2017-08-18 | 2017-12-15 | 四川普莱美生物科技集团有限公司 | Modeling method, preparation, the preparation method of preparation, rhesus macaque pulmonary fibrosis model and its application of rhesus macaque pulmonary fibrosis model |
| CN112274294A (en) * | 2020-10-14 | 2021-01-29 | 苏州西山中科药物研究开发有限公司 | Method for inducing lung injury model of experimental animal with different severity by intratracheal spraying of different doses of modeling drug |
| CN113100177A (en) * | 2021-06-16 | 2021-07-13 | 澎立生物医药技术（上海）有限公司 | Bleomycin-induced pulmonary fibrosis cynomolgus monkey model and application thereof |
| CN113287569A (en) * | 2021-05-27 | 2021-08-24 | 四川康城生物科技有限公司 | Construction method of animal model with low immunity |
| WO2022094929A1 (en) * | 2020-11-06 | 2022-05-12 | 四川大学华西医院 | Method for establishing animal acute lung injury model |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104784676A (en) * | 2015-05-06 | 2015-07-22 | 中国科学院合肥物质科学研究院 | Method for establishing rat single lung pulmonary fibrosis model |
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| CN104784676A (en) * | 2015-05-06 | 2015-07-22 | 中国科学院合肥物质科学研究院 | Method for establishing rat single lung pulmonary fibrosis model |
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| 李伟峰等: "气管内滴入与雾化博莱霉素致小鼠肺纤维化模型的比较研究", 《南方医科大学学报》 * |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107469069A (en) * | 2017-08-18 | 2017-12-15 | 四川普莱美生物科技集团有限公司 | Modeling method, preparation, the preparation method of preparation, rhesus macaque pulmonary fibrosis model and its application of rhesus macaque pulmonary fibrosis model |
| CN112274294A (en) * | 2020-10-14 | 2021-01-29 | 苏州西山中科药物研究开发有限公司 | Method for inducing lung injury model of experimental animal with different severity by intratracheal spraying of different doses of modeling drug |
| WO2022094929A1 (en) * | 2020-11-06 | 2022-05-12 | 四川大学华西医院 | Method for establishing animal acute lung injury model |
| CN113287569A (en) * | 2021-05-27 | 2021-08-24 | 四川康城生物科技有限公司 | Construction method of animal model with low immunity |
| CN113100177A (en) * | 2021-06-16 | 2021-07-13 | 澎立生物医药技术（上海）有限公司 | Bleomycin-induced pulmonary fibrosis cynomolgus monkey model and application thereof |
| CN113100177B (en) * | 2021-06-16 | 2022-02-22 | 澎立生物医药技术（上海）有限公司 | Bleomycin-induced pulmonary fibrosis cynomolgus monkey model and application thereof |
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| CN105126075B (en) | 2018-12-11 |
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