CN115414346B - Method for constructing pulmonary fibrosis model by using amiodarone - Google Patents
Method for constructing pulmonary fibrosis model by using amiodarone Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
- A61K31/343—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/027—New breeds of vertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/14—Peptides containing saccharide radicals; Derivatives thereof, e.g. bleomycin, phleomycin, muramylpeptides or vancomycin
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
Abstract
The invention discloses a method for constructing a pulmonary fibrosis model by using amiodarone, which is characterized by comprising the following steps of: the construction method comprises the following steps: the first step: preparing a proper amount of mice, and adaptively raising the mice; and a second step of: the bleomycin is administered by intraperitoneal injection, intravenous injection, subcutaneous injection, intratracheal instillation or aerosol inhalation, and the bleomycin is induced by single-dose administration or multiple-dose administration; and a third step of: placing all the mice at 18-20 ℃ and with relative humidity of 53-58%, and feeding normally in a day-night alternate environment, and the fourth step: by aerosol inhalation of amiodarone to the mice induced in the second step. The pulmonary fibrosis model prepared by the invention has the characteristics of short time for inducing pulmonary fibrosis, uniform pulmonary fibrosis distribution, small death rate of test animals, high success rate of the model, good stability of the model and low cost, and overcomes the uneven pulmonary fibrosis distribution and the self-healing tendency of the single bleomycin model.
Description
Technical Field
The invention relates to the technical field of amiodarone application, in particular to a method for constructing a pulmonary fibrosis model by using amiodarone.
Background
Amiodarone is an antiarrhythmic drug, which is widely used in clinic and is mainly used for treating ventricular arrhythmia and atrial fibrillation, and the side effect is pulmonary fibrosis with unknown mechanism of action. The induction of pulmonary fibrosis with amiodarone at the molecular level has been widely reported, but there is no report of using amiodarone for an animal model of pulmonary fibrosis.
With the aggravation of domestic air pollution degree, particles in pollutants directly enter the lung, cause slow and continuous damage to alveoli and vascular endothelium, and induce the generation of pulmonary fibrosis. Pulmonary fibrosis is a diffuse lung disease of unknown cause that ultimately leads to severe destruction of the structure and function of the lung. Since the pathogenesis of pulmonary fibrosis is not yet clear, exploring an animal model of pulmonary fibrosis is a key to developing therapeutic drugs.
The animal model for pulmonary fibrosis must have histological features capable of replicating typical common interstitial pneumonia, and the animal model for pulmonary fibrosis has the characteristics of persistence and irreversibility, low economic cost, repeatable model and short molding time. There are a number of animals currently available as model vectors for testing lung fibrosis, including mice, rats, hamsters, rabbits, dogs, and non-human primates, but the most commonly used are C57BL/6 mice.
Heretofore, there are many methods for inducing pulmonary fibrosis in experimental animals, including chemical methods (bleomycin, silica, fluorescein isothiocyanate), radiation, and gene induction methods (transduction of specific gene expression using transgenic systems or viral vectors), and the like. However, each method has defects and shortages, and is mainly limited in that the induced pulmonary fibrosis model lacks the histological features of common interstitial pneumonia and the method is technically and economically expensive, so that the method cannot be popularized and used. For example, the lung fibrosis model induced by methods such as silica aerosol inhalation, fluorescein isothiocyanate intratracheal administration, radiation irradiation and the like lacks the characteristic of common interstitial pneumonia, and the silica aerosol inhalation model, the transgenic system or the virus vector transduction special gene expression model have high technical requirements and high economic cost.
Bleomycin is a most commonly used drug for inducing pulmonary fibrosis at present, and has relatively low price, convenient acquisition route and strong repeatability. C57BL/6 mice are most sensitive to bleomycin and are the most commonly used lung fibrosis experimental animals at present. The routes of administration of bleomycin include intraperitoneal injection, intravenous injection, subcutaneous injection and intratracheal instillation, with intratracheal instillation being most commonly used at present, with single dose instillation being most commonly used. However, it has been studied that single intratracheal instillation of bleomycin drug is not uniformly distributed, lacks typical characteristic of ordinary interstitial pneumonia, focus is mainly distributed around the phylum and bronchi, is different from the focus of idiopathic pulmonary interstitial fibrosis which is mainly distributed under the pleura, has poor model stability, and is different from the course of progressive deterioration of idiopathic pulmonary interstitial fibrosis, multiple intratracheal instillation of bleomycin pulmonary fibrosis model replicates many pathological features of idiopathic pulmonary interstitial fibrosis (spatial heterogeneity of pulmonary fibrosis, transformation of epithelial cells and hyperplasia of alveolar epithelial cells), and has good model stability, but takes very long time for modeling (16 weeks), and easily causes death of experimental animals. The tail vein injection and the intraperitoneal injection of bleomycin belong to systemic administration, and the focus of bleomycin is mainly distributed under the pleura and around the blood vessels, and is similar to the pathological changes of idiopathic pulmonary interstitial fibrosis. The single dose tail vein injection bleomycin model has insufficient stability, and the pulmonary fibrosis has obvious self-limitation after 6 weeks of administration. The model of the bleomycin injected by tail vein for many times has better stability, but the modeling takes 6 weeks, and the repeated bleomycin injected by tail vein has high technical requirement and slightly poorer operability.
Disclosure of Invention
The invention aims to provide a method for constructing a pulmonary fibrosis model by using amiodarone, which aims to solve the problem of rapidly and stably establishing and stabilizing a continuous pulmonary fibrosis animal model.
In order to achieve the above purpose, the present invention provides the following technical solutions: a method of constructing a model of pulmonary fibrosis using amiodarone, the construction method comprising:
the first step: preparing a proper amount of mice, and adaptively raising the mice;
and a second step of: the bleomycin is administered by intraperitoneal injection, intravenous injection, subcutaneous injection, intratracheal instillation or aerosol inhalation, and the bleomycin is induced by single-dose administration or multiple-dose administration;
and a third step of: all the mice are placed at 18-20 ℃ and have a relative humidity of 53-58% and are fed normally in a day-night alternating environment.
Fourth step: by aerosol inhalation of amiodarone to the mice induced in the second step, the amount of amiodarone inhaled by the mice is 2.5-10mg/kg.
The mice induced by using Bo Lei Meisu are laboratory rats and mice.
The mice include C57BL/6 mice.
The mode of aerosol inhalation of amiodarone comprises the use of an aerosol box for aerosol and insertion for aerosol after anesthesia.
Preferably, the method for atomizing and inhaling amiodarone after anesthesia is to use an animal mirror to press down the mouse tongue root to expose glottis, and the atomizer injects amiodarone from the glottis insertion organ.
Compared with the prior art, the invention has the beneficial effects that:
1. the pulmonary fibrosis model prepared by the invention has the characteristics of short time for inducing pulmonary fibrosis, uniform pulmonary fibrosis distribution, small death rate of test animals, high success rate of the model, good stability of the model and low cost, and overcomes the uneven pulmonary fibrosis distribution and the self-healing tendency of the single bleomycin model.
Drawings
FIG. 1 is an alveoli score of the present invention;
FIG. 2 is a graph showing the lung fibrosis score according to the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example I
Referring to fig. 1 and 2, a method for constructing a lung fibrosis model using amiodarone is provided, the construction method being as follows:
the first step: preparing a proper amount of mice, and adaptively raising the mice;
and a second step of: the bleomycin is administered by intraperitoneal injection, intravenous injection, subcutaneous injection, intratracheal instillation or aerosol inhalation, and the bleomycin is induced by single-dose administration or multiple-dose administration;
and a third step of: all the mice are placed at 18-20 ℃ and have a relative humidity of 53-58% and are fed normally in a day-night alternating environment.
Fourth step: by aerosol inhalation of amiodarone to the mice induced in the second step, the amount of amiodarone inhaled by the mice is 2.5-10mg/kg.
Mice induced with blog Lei Meisu are laboratory rats and mice.
Mice include C57BL/6 mice.
Ways of nebulizing the inhaled amiodarone include nebulization using a nebulizing cartridge and insertion nebulization after anesthesia.
The method for atomizing and inhaling amiodarone after anesthesia is to use an animal mirror to press down the mouse tongue root to expose glottis, and the atomizer is inserted into organs from the glottis to inject amiodarone.
The amount of amiodarone inhaled by murine aerosol is 2.5-10mg/kg.
The bleomycin induction method of the above method further comprises aerosolizing the inhaled bleomycin.
When the embodiment is used, the following steps are adopted: (1) grouping of animals: 8-year-old C57BL/6 mice, male and female, were randomly divided into model groups and control groups, and each group was subjected to adaptive breeding for 7d.
(2) All mice are anesthetized by intraperitoneal injection of chloral hydrate, the trachea is separated after the anesthesia is effective, a proper amount of bleomycin (5 mg/kg) is dripped into the trachea, and the mice are erected and vertically rotated after dripping, so that the medicine is uniformly distributed in the lung.
(3) All mice were placed at 18-20deg.C with relative humidity of 53-58% and fed normally in a day-night alternating environment.
(4) All mice were nebulized using a mouse nebulizing cartridge, wherein 5mg/kg amiodarone was nebulized by the model group, once daily, physiological saline was inhaled by the control group, once daily, and samples were obtained by batch sacrifice of mice at 2, 4, 6, 8, 10 weeks, respectively.
Example II
Referring to fig. 1 and 2, this embodiment further illustrates a method for constructing a pulmonary fibrosis model using amiodarone, the method of construction being as follows:
the first step: preparing a proper amount of mice, and adaptively raising the mice;
and a second step of: the bleomycin is administered by intraperitoneal injection, intravenous injection, subcutaneous injection, intratracheal instillation or aerosol inhalation, and the bleomycin is induced by single-dose administration or multiple-dose administration;
and a third step of: all the mice are placed at 18-20 ℃ and have a relative humidity of 53-58% and are fed normally in a day-night alternating environment.
Fourth step: by aerosol inhalation of amiodarone to the mice induced in the second step, the amount of amiodarone inhaled by the mice is 2.5-10mg/kg.
Mice induced with blog Lei Meisu are laboratory rats and mice.
Mice include C57BL/6 mice.
Ways of nebulizing the inhaled amiodarone include nebulization using a nebulizing cartridge and insertion nebulization after anesthesia.
The method for atomizing and inhaling amiodarone after anesthesia is to use an animal mirror to press down the mouse tongue root to expose glottis, and the atomizer is inserted into organs from the glottis to inject amiodarone.
The amount of amiodarone inhaled by murine aerosol is 2.5-10mg/kg.
The bleomycin induction method of the above method further comprises aerosolizing the inhaled bleomycin.
When the embodiment is used, the following steps are adopted: lung tissue distribution of evans blue: the Evansi blue solution is atomized into the trachea of the mice, sacrificed 5min after administration, and the whole lung is separated, so that the lungs of the mice in the control group are unevenly distributed in blocks, the Evansi blue in the lungs of the mice in the model group are evenly distributed, and the Evansi blue in the lungs of the mice in the model group are scattered.
Example III
Referring to fig. 1 and 2, this embodiment further illustrates, for other examples, a method for constructing a lung fibrosis model using amiodarone, the construction method being as follows:
the first step: preparing a proper amount of mice, and adaptively raising the mice;
and a second step of: the bleomycin is administered by intraperitoneal injection, intravenous injection, subcutaneous injection, intratracheal instillation or aerosol inhalation, and the bleomycin is induced by single-dose administration or multiple-dose administration;
and a third step of: all the mice are placed at 18-20 ℃ and have a relative humidity of 53-58% and are fed normally in a day-night alternating environment.
Fourth step: by aerosol inhalation of amiodarone to the mice induced in the second step, the amount of amiodarone inhaled by the mice is 2.5-10mg/kg.
Mice induced with blog Lei Meisu are laboratory rats and mice.
Mice include C57BL/6 mice.
Ways of nebulizing the inhaled amiodarone include nebulization using a nebulizing cartridge and insertion nebulization after anesthesia.
The method for atomizing and inhaling amiodarone after anesthesia is to use an animal mirror to press down the mouse tongue root to expose glottis, and the atomizer is inserted into organs from the glottis to inject amiodarone.
The amount of amiodarone inhaled by murine aerosol is 2.5-10mg/kg.
The bleomycin induction method of the above method further comprises aerosolizing the inhaled bleomycin.
When the embodiment is used, the following steps are adopted: alveolitis and pulmonary fibrosis scores: the pulmonary alveoli inflammation and pulmonary fibrosis inflammation levels are higher after 2 weeks of model assembly, the peak value is reached after 4 weeks of model assembly, and the decrease is not obvious after 10 weeks of model assembly; alveolar inflammation and pulmonary fibrosis inflammation peak 4-6 weeks after molding of the control group, and decrease significantly from 8 weeks to the lowest after molding, and decrease to the lowest after 10 weeks. (alveolar inflammatory degree score. 0. No alveolitis, lung tissue was normal; 1 mild alveolitis, clear mononuclear cells, thickened alveolar spaces, lesions limited to subpleura and a lesion range <20%, 2 moderate alveolitis, inflammatory lesions mainly located under pleura and a lesion range of 20% -50%, 3 severe alveolitis, diffuse inflammatory lesions and a lesion range > 50%) (lung fibrosis degree score. 0 is free of fibrosis, normal alveolar structure, 1 dispersed in mild fibrosis foci, alveolar space thickness less than or equal to 3 times normal value, partial alveolar space enlarged, alveolar wall thinned but free of mass pulmonary fibrosis, 2 obvious pulmonary fibrosis change, alveolar space thickness >3 times normal value and accompanied by rope knot-like change, but unfused, partial alveolar space enlarged, alveolar wall thinned but free of mass pulmonary fibrosis, 3 minutes continuous pulmonary fibrosis, alveolar space thickness >3 times normal value, partial alveolar space enlarged, alveolar wall thinned, 4 minutes single mass pulmonary fibrosis foci, focal range 10% -50%, lung morphology structure still can be thin and visible, 6 minutes of large sheets of pulmonary fibrosis is almost lost, 7% of large mass fibrosis is almost lost, and at least 7% of large mass of pulmonary structure is almost lost, and at least 5% of fibrous mass is almost lost, and the fibrous structure of the large alveolar space is almost lost.
It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (2)
1. A method of constructing a model of pulmonary fibrosis using amiodarone, comprising: the construction method comprises the following steps:
the first step: preparing a proper amount of mice, and adaptively raising the mice;
and a second step of: the bleomycin is administered by intraperitoneal injection, intravenous injection, subcutaneous injection, intratracheal instillation or aerosol inhalation, and the bleomycin is induced by single-dose administration or multiple-dose administration;
and a third step of: placing all the mice at 18-20 ℃ and with relative humidity of 53-58%, and feeding normally in a day-night alternating environment;
fourth step: carrying out atomized inhalation of amiodarone on the mice induced in the second step, wherein the amount of amiodarone inhaled by the mice is 2.5-10mg/kg;
the bleomycin-induced mice refer to laboratory rats and mice;
the mice include C57BL/6 mice;
the mode of aerosol inhalation of amiodarone comprises the use of an aerosol box for aerosol and insertion for aerosol after anesthesia.
2. A method of constructing a model of pulmonary fibrosis using amiodarone according to claim 1 wherein: the method for atomizing and inhaling amiodarone after anesthesia is to use an animal mirror to press down the mouse tongue root to expose glottis after the mouse is anesthetized, and the amiodarone is injected from a glottis insertion organ by an atomizer.
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| CN202211041855.XA CN115414346B (en) | 2022-08-29 | 2022-08-29 | Method for constructing pulmonary fibrosis model by using amiodarone |
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| CN202211041855.XA CN115414346B (en) | 2022-08-29 | 2022-08-29 | Method for constructing pulmonary fibrosis model by using amiodarone |
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| CN115414346B true CN115414346B (en) | 2023-09-29 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101642451A (en) * | 2008-08-08 | 2010-02-10 | 上海安普生物科技有限公司 | New application of amiodarone and derivatives thereof |
| WO2017103108A1 (en) * | 2015-12-16 | 2017-06-22 | Singapore Health Services Pte Ltd | Treatment of fibrosis |
| CN107115343A (en) * | 2017-05-16 | 2017-09-01 | 温州市人民医院 | A kind of Rat Pulmonary fibrosis model |
| CN108042239A (en) * | 2017-12-22 | 2018-05-18 | 上海市胸科医院 | A kind of method for building up of noninvasive pulmonary fibrosis model |
| CN109836423A (en) * | 2019-04-03 | 2019-06-04 | 黑龙江中医药大学 | Noval chemical compound, the preparation method and its usage of one kind prevention or treatment pulmonary fibrosis disease |
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| US20090131359A1 (en) * | 2007-11-19 | 2009-05-21 | Atamas Sergei P | Antifibrotic therapy |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101642451A (en) * | 2008-08-08 | 2010-02-10 | 上海安普生物科技有限公司 | New application of amiodarone and derivatives thereof |
| WO2017103108A1 (en) * | 2015-12-16 | 2017-06-22 | Singapore Health Services Pte Ltd | Treatment of fibrosis |
| CN107115343A (en) * | 2017-05-16 | 2017-09-01 | 温州市人民医院 | A kind of Rat Pulmonary fibrosis model |
| CN108042239A (en) * | 2017-12-22 | 2018-05-18 | 上海市胸科医院 | A kind of method for building up of noninvasive pulmonary fibrosis model |
| CN109836423A (en) * | 2019-04-03 | 2019-06-04 | 黑龙江中医药大学 | Noval chemical compound, the preparation method and its usage of one kind prevention or treatment pulmonary fibrosis disease |
Non-Patent Citations (2)
| Title |
|---|
| "Animal models of drug‑induced pulmonary fibrosis: an overview of molecular mechanisms and characteristics";Shuchan Li et al.;《Cell Biol Toxicol》;第38卷;第699–723页 * |
| "三种方法气管灌注博来霉素诱导小鼠 肺纤维化的比较";王志超等;《中国比较医学杂志》;第29卷(第5期);第51-57页 * |
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Denomination of invention: A method of constructing a pulmonary fibrosis model using amiodarone Granted publication date: 20230929 Pledgee: Agricultural Bank of China Xiangtan County Branch Pledgor: Hunan Furui Biomedical Technology Co.,Ltd. Registration number: Y2024980000042 |