CN107349225A - A kind of experimental mouse model and construction method of caprophyl transplanting - Google Patents
A kind of experimental mouse model and construction method of caprophyl transplanting Download PDFInfo
- Publication number
- CN107349225A CN107349225A CN201710549233.0A CN201710549233A CN107349225A CN 107349225 A CN107349225 A CN 107349225A CN 201710549233 A CN201710549233 A CN 201710549233A CN 107349225 A CN107349225 A CN 107349225A
- Authority
- CN
- China
- Prior art keywords
- mouse
- solution
- construction method
- caprophyl
- cathartic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
-
- 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
- A61K31/74—Synthetic polymeric materials
- A61K31/765—Polymers containing oxygen
- A61K31/77—Polymers containing oxygen of oxiranes
-
- 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
Abstract
The invention discloses the experimental mouse model and construction method of a kind of caprophyl transplanting, the construction method comprises the following steps：Gavage injection of antibiotics solution is carried out to mouse, the number of daily gavage injection is at least 1 time, and the number of days of gavage injection is at least 1；After mouse stomach injection of antibiotics solution, drunk using cathartic solution as water source, the number of days for drinking cathartic solution is at least 1；After mouse drinks cathartic solution, caprophyl transplanting is carried out to mouse, that is, builds the experimental mouse model of caprophyl transplanting.The construction method of experimental mouse model of the present invention is simple, and can true and reliable response situation.
Description
Technical field
The invention belongs to biological technical field, more particularly to the experimental mouse model and construction method of a kind of transplanting of caprophyl.
Background technology
Recently, modern medicine institute has been becoming increasingly recognized that important function of the enteric bacteria in human body, will maintain or again
Build critical treatment means of the human body intestinal canal normal flora as a variety of diseases.As enteron aisle and the treatment means of parenterally disease,
Caprophyl transplanting (Fecal Microbiota Transplantation, FMT) is to transplant the functional flora in people's health excrement
Into patient's gastrointestinal tract, new gut flora is rebuild.Caprophyl transplanting is gradually paid attention to, but it comes with some shortcomings, and having must
Experimental mouse model is built to be furtherd investigate and analyzed.
At present, the Germ-free mouse model of caprophyl transplantation experiments is not only built cumbersome, and modelling effect is poor：It is difficult to true anti-
Answer situation.
The content of the invention
A kind of experiment of caprophyl transplanting is provided it is an object of the invention to overcome the shortcomings of the prior art part
Mouse model and its construction method, the construction method of experimental mouse model of the present invention is simple, and can true and reliable response situation.
The technical solution adopted by the present invention is：A kind of construction method of the experimental mouse model of caprophyl transplanting, including it is following
Step：After being handled using antibiotic and cathartic mouse, caprophyl transplanting is carried out to mouse, obtains the experiment mice mould of caprophyl transplanting
Type.
Gavage injection of antibiotics solution is carried out to mouse, itself a part of gut flora can be removed；Then, mouse, which is drunk, rushes down
Drug solns, it can further remove itself gut flora；Of the same race or xenogenesis caprophyl donor bacterium transplanting is carried out afterwards, and donor bacterium can
The structure of spontaneous flora is significantly changed, and is colonized in acceptor enteron aisle inside points, constructs the experimental mouse model of caprophyl transplanting.
As the improvement of above-mentioned technical proposal, described construction method comprises the following steps：
1) gavage injection of antibiotics solution is carried out to mouse, the number of daily gavage injection is at least 1 time, gavage injection
Number of days is at least 1；
2) after mouse stomach injection of antibiotics solution, drunk using cathartic solution as water source, drink the day of cathartic solution
Number is at least 1；
3) after mouse drinks cathartic solution, caprophyl transplanting is carried out to mouse, that is, constructs the experiment mice mould of caprophyl transplanting
Type.
As the improvement of above-mentioned technical proposal, the mouse is SPF (Specific Pathogene Free, without specific disease
Opportunistic pathogen) mouse.
The animal model of caprophyl transplanting has the following disadvantages frequently with germfree mouse, germfree mouse when building model：1)
Environmental condition requires high, and cost is high；2) breeding rate is low, and some gene defection types even can not be raised and bred；3) vitality is poor,
The death rate is high after disease modeling；4) congenital immunity and neurologically handicapped be present, cause relevant disease result of study unreliable.Use enteron aisle
The SPF mouse of cleaning replace the model of germfree mouse structure caprophyl transplanting, can simplify experiment, less with experimental animal, reduce into
This；SPF level mouse approximation healthy mices, more truly reflect experimental result than germfree mouse.
As the improvement of above-mentioned technical proposal, the antibiotic solution is vancomycin normal saline solution, the cathartic
Solution is polyalkylene glycol compound normal saline solution.
Further improved as above-mentioned technical proposal, vancomycin concentration is in the vancomycin normal saline solution
20mg/ml, polyalkylene glycol compound concentration is 200mg/ml in polyalkylene glycol compound normal saline solution.
Further improved as above-mentioned technical proposal, it is water-soluble that the daily gavage of mouse injects 2 vancomycin physiology salts
Liquid, the vancomycin normal saline solution of each gavage injection is 0.25ml, and gavage injection fills vancomycin life to mouse for three days on end
Manage saline solution；Mouse drinks polyalkylene glycol compound normal saline solution for three days on end.
In addition, the present invention also carries a kind of experimental mouse model of the caprophyl transplanting of the construction method structure described in use.
In addition, the present invention also provides purposes of the SPF mouse in the experimental mouse model in structure caprophyl transplanting.
In addition, the present invention also offer antibiotic and cathartic are used in combination in the experimental mouse model of structure caprophyl transplanting
Purposes.
Beneficial effects of the present invention are：The present invention provides a kind of experimental mouse model and construction method of caprophyl transplanting, this
The construction method of invention experimental mouse model is：First to mouse stomach injection of antibiotics solution, then allow mouse using cathartic solution as
Water source is drunk, and finally carries out caprophyl transplanting, construction method relatively easyization；Germfree mouse is replaced with SPF mouse, is not only dropped
Low cost, and the experimental mouse model constructed is stablized, worked well relatively：Can true and reliable response situation.
Brief description of the drawings
Fig. 1 is the Colon and rectum pathological examination figure of the experimental mouse model of caprophyl transplanting；
Fig. 2 is the gene sequencing figure of gut flora during the caecum of the experimental mouse model of caprophyl transplanting is defecated；
In Fig. 1 and Fig. 2, FMT is the experimental mouse model of caprophyl transplanting.
Embodiment
To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with specific embodiments and the drawings pair
The present invention is described further.
The structure of the experimental mouse model of the caprophyl of embodiment 1 transplanting
1) the daily gavage of mouse injects 2 vancomycin normal saline solutions, and gavage is injected for three days on end；Each gavage note
The vancomycin normal saline solution penetrated is 0.25ml, and vancomycin concentration is 20mg/ml in vancomycin normal saline solution；
2) after mouse stomach injection vancomycin normal saline solution, using polyalkylene glycol compound normal saline solution as water source
Drunk, mouse is drunk for three days on end；
3) after mouse drinks polyalkylene glycol compound normal saline solution, excrement is carried out to mouse with the gut flora of stroke patient
Bacterium is transplanted, that is, constructs the experimental mouse model of caprophyl transplanting.
Embodiment 2
The experimental mouse model transplanted using caprophyl in embodiment 1 is object (setting one group of blank control), to Colon and rectum stage casing
Tissue carries out pathological examination, and pathological examination comprises the following steps：
1) specimen sampling：Mouse fasting 12h, free water before collection of specimens.Take the de- white execution of mouse cervical vertebra, 75% ethanol
3min is soaked, mouse is taken out and is placed on sterile gauze, dorsal position.Through mouse median abdominal incision, skin is cut off, exposure stomach wall, is carried
Play peritonaeum and cut off.Exposure abdominal tissue, careful point of whole intestinal tubes from ileocecus to tripe door, with ice normal saline flushing
Enteric cavity, intestinal contents is washed away, leave and take colon and fixed with 10% formalin；
2) paraffin embedded tissues make
Dehydration：70% ethanol is stayed overnight, 80% ethanol 60min, 90% ethanol 60min, 95% ethanol 30min, 95% ethanol
30min, 100% ethanol 15min, 100% ethanol 15min；It is transparent：Dimethylbenzene 15min, dimethylbenzene 15min；Soak vinegar：Paraffin
30min, paraffin 90min；Embedding；
3) haematoxylin Yihong (HE) staining procedure
The tissue block section of FFPE, slice thickness are 5 μm；42 DEG C of water tanks open up piece, with anti-after histotomy flattening
De- slide takes piece, 65 DEG C of insulating box bakings is placed in, to reduce flake；Paraffin section routinely dewaxes, aquation：Dimethylbenzene 10min,
Dimethylbenzene 5min, 100% ethanol 5min, 100% ethanol 5min, 95% ethanol 3min, 80% ethanol 3min, 70% ethanol
3min, circulating water rinse 5min, 1~3min of haematoxylin dyeing, and flowing water rinses 5min, 1% hydrochloride alcohol differentiation 5s, and flowing water rinses
5s, 0.5% light ammoniacal liquor, 30~45s of anti-indigo plant, flowing water rinse 5min, eosin stains 1min, and flowing water, which dips in, to be washed under 3~5；Graded ethanol takes off
Water：70% ethanol, which dips in, to be washed under 3-5, and 80% ethanol, which dips in, to be washed under 3-5, and 95% ethanol, which dips in, to be washed under 3-5,100% ethanol 10min, and 100%
Ethanol 10min；It is transparent：The 10min of dimethylbenzene I, the 10min of dimethylbenzene II, neutral gum mounting, microscopy.
Experimental result is as shown in figure 1, find that pathological change occurs for the enteron aisle of the experiment mice of caprophyl transplanting：Absorb cell
Structure disturbance, and with partial exfoliation；The mucous layer and visible lymphocyte of submucosa, neutrophil leucocyte and plasmacytosis,
Infiltration；Submucosa part thickens and with lymphocytic infiltration.Thus, it can be seen that the experiment mice of caprophyl transplanting of the invention
Model successfully constructs.
Embodiment 3
The experimental mouse model transplanted using caprophyl in embodiment 1 is object (setting one group of blank control), in being defecated to caecum
Gut flora carries out the detection of 16S rRNA gene sequencing.Experimental result is as shown in Fig. 2 find the experiment mice that caprophyl is transplanted
Obvious change occurs for gut flora.As can be seen here, the experimental mouse model of caprophyl of the invention transplanting successfully constructs.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than the present invention is protected
The limitation of scope is protected, although being explained in detail with reference to preferred embodiment to the present invention, one of ordinary skill in the art should
Understand, technical scheme can be modified or equivalent substitution, without departing from the essence of technical solution of the present invention
And scope.
Claims (9)
1. a kind of construction method of the experimental mouse model of caprophyl transplanting, it is characterised in that comprise the following steps：
After being handled using antibiotic and cathartic mouse, caprophyl transplanting is carried out to mouse, obtains the experiment mice mould of caprophyl transplanting
Type.
2. as profit requires 1 construction method, it is characterised in that：Including step in detail below：
1) gavage injection of antibiotics solution is carried out to mouse, the number of daily gavage injection is at least 1 time, the number of days of gavage injection
At least 1 day；
2) after mouse stomach injection of antibiotics solution, drunk using cathartic solution as water source, drink the number of days of cathartic solution extremely
It is 1 less；
3) after mouse drinks cathartic solution, caprophyl transplanting is carried out to mouse, that is, constructs the experimental mouse model of caprophyl transplanting.
3. construction method as claimed in claim 1 or 2, it is characterised in that：The mouse is SPF mouse.
4. construction method as claimed in claim 1 or 2, it is characterised in that：The antibiotic solution is vancomycin physiology salt
The aqueous solution, the cathartic solution are polyalkylene glycol compound normal saline solution.
5. construction method as claimed in claim 4, it is characterised in that：Vancomycin in the vancomycin normal saline solution
Concentration is 20mg/ml, and polyalkylene glycol compound concentration is 200mg/ml in polyalkylene glycol compound normal saline solution.
6. construction method as claimed in claim 5, it is characterised in that：The daily gavage of mouse injects 2 vancomycin physiology salts
The aqueous solution, the vancomycin normal saline solution of each gavage injection is 0.25ml, and gavage injection fills mould through the ages to mouse for three days on end
Plain normal saline solution；Mouse drinks polyalkylene glycol compound normal saline solution for three days on end.
A kind of 7. experimental mouse model of the caprophyl transplanting of construction method structure using as described in any one of claim 1~6.
Purposes of the 8.SPF mouse in the experimental mouse model in structure caprophyl transplanting.
9. the purposes being used in combination in the experimental mouse model of structure caprophyl transplanting of antibiotic and cathartic.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710549233.0A CN107349225A (en) | 2017-07-06 | 2017-07-06 | A kind of experimental mouse model and construction method of caprophyl transplanting |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710549233.0A CN107349225A (en) | 2017-07-06 | 2017-07-06 | A kind of experimental mouse model and construction method of caprophyl transplanting |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107349225A true CN107349225A (en) | 2017-11-17 |
Family
ID=60291726
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710549233.0A Pending CN107349225A (en) | 2017-07-06 | 2017-07-06 | A kind of experimental mouse model and construction method of caprophyl transplanting |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107349225A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110122412A (en) * | 2019-05-21 | 2019-08-16 | 中国科学院合肥物质科学研究院 | A kind of construction method of lung's flora transplanting rodent model |
| CN110122413A (en) * | 2019-05-21 | 2019-08-16 | 中国科学院合肥物质科学研究院 | A kind of construction method of lung's flora rodent model |
| CN110178787A (en) * | 2019-05-08 | 2019-08-30 | 河南中医药大学 | A method of self-closing disease rat model is established with caprophyl grafting |
| CN114606327A (en) * | 2022-01-17 | 2022-06-10 | 华南理工大学 | Method for evaluating using mode of antibiotics |
| CN115281154A (en) * | 2022-08-30 | 2022-11-04 | 中国人民解放军联勤保障部队第九〇四医院 | Construction method of pseudo-sterile rat model |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103446576A (en) * | 2013-09-12 | 2013-12-18 | 光明乳业股份有限公司 | Modeling method of helicobacter pylori infection animal model |
| CN104739861A (en) * | 2015-04-09 | 2015-07-01 | 重庆医科大学 | Construction method of 'humanized' microorganism disorder depression model |
| CN105942511A (en) * | 2016-05-26 | 2016-09-21 | 惠州同富康生物科技有限公司 | Green-plum-contained health care food composition capable of regulating intestinal flora, and preparation method |
-
2017
- 2017-07-06 CN CN201710549233.0A patent/CN107349225A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103446576A (en) * | 2013-09-12 | 2013-12-18 | 光明乳业股份有限公司 | Modeling method of helicobacter pylori infection animal model |
| CN104739861A (en) * | 2015-04-09 | 2015-07-01 | 重庆医科大学 | Construction method of 'humanized' microorganism disorder depression model |
| CN105942511A (en) * | 2016-05-26 | 2016-09-21 | 惠州同富康生物科技有限公司 | Green-plum-contained health care food composition capable of regulating intestinal flora, and preparation method |
Non-Patent Citations (3)
| Title |
|---|
| 张发明: "粪菌移植的概念、历史、现状和未来", 《中国内镜杂志》 * |
| 王瑞君: "抗生素诱导的菌群失调模型的建立及其应用于培菲康疗效的评价", 《中国硕士学位论文 医药卫生科技辑》 * |
| 肖英平: "金华猪和长白猪粪便微生物移植小鼠的肠道古菌结构分析", 《动物营养学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110178787A (en) * | 2019-05-08 | 2019-08-30 | 河南中医药大学 | A method of self-closing disease rat model is established with caprophyl grafting |
| CN110122412A (en) * | 2019-05-21 | 2019-08-16 | 中国科学院合肥物质科学研究院 | A kind of construction method of lung's flora transplanting rodent model |
| CN110122413A (en) * | 2019-05-21 | 2019-08-16 | 中国科学院合肥物质科学研究院 | A kind of construction method of lung's flora rodent model |
| CN114606327A (en) * | 2022-01-17 | 2022-06-10 | 华南理工大学 | Method for evaluating using mode of antibiotics |
| CN115281154A (en) * | 2022-08-30 | 2022-11-04 | 中国人民解放军联勤保障部队第九〇四医院 | Construction method of pseudo-sterile rat model |
| CN115281154B (en) * | 2022-08-30 | 2023-11-03 | 中国人民解放军联勤保障部队第九〇四医院 | Method for constructing pseudo-sterile rat model |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107349225A (en) | A kind of experimental mouse model and construction method of caprophyl transplanting | |
| CN102271722B (en) | Cell-scaffold constructs | |
| CN204106683U (en) | A kind of peritoneal irrigation drain sepage management devices | |
| CN104546868A (en) | Establishing method of primary generation humanized mammary cancer transplantation tumor model based on Chinese population and model | |
| Chen et al. | Preparation of decellularized liver scaffolds and recellularized liver grafts | |
| CN111903606A (en) | Construction method of lung cancer in-situ PDX model inoculated by percutaneous puncture | |
| CN101305710B (en) | Corpus antiseptic and its preparation method | |
| Felgendreff et al. | Identification of tissue sections from decellularized liver scaffolds for repopulation experiments | |
| CN102552935B (en) | Use of hepatocyte nuclear factor-1alpha in treatment of chronic liver disease | |
| CN108079363A (en) | A kind of kit and its application that cell processing is taken off for animal tissue | |
| WO2019029084A1 (en) | 3d printed artificial endometrium and preparation method and application thereof | |
| CN105266916A (en) | Extracted tooth processing and storage method for reutilization | |
| Yari et al. | Anatomical and histological study of the excretory system in the Bosc’s Fringe-Toed Lizard (Acanthodactylus boskianus) | |
| CN105555230A (en) | Base material for forming connective tissue structure and method for producing connective tissue structure | |
| CN108030914A (en) | A kind of Matrigel raw material and preparation method and application | |
| CN107296007B (en) | Construction method of animal model of metacercaria pulmonary cyst disease of sandrat afternoon | |
| Jabbar et al. | Anatomical And Histological Invistgation Of The Kidney In Goat (Capra hircus) | |
| CN108142342A (en) | A kind of method that crab class individual mark is carried out using electronic tag | |
| CN1920008A (en) | Establishing method of rat primary liver cancer model | |
| Spadola et al. | Hernioplasty with Peritoneal Flap for the Surgical Treatment of Umbilical Hernia in Swine | |
| CN101926307A (en) | Establishment method of animal model in mice infected with sparganosis mansoni | |
| CN106176747B (en) | Tacrine is preparing the application in treating treating echinococcosis | |
| Clark et al. | Anatomy of an agricultural antagonist: Feeding complex structure and function of three xylem sap‐feeding insects illuminated with synchrotron‐based 3D imaging | |
| CN115500317B (en) | Method for establishing mouse orthotopic bladder cancer model | |
| CN107970274A (en) | Application of the pueraria lobata in terms of mesenchymal stem cell migration is promoted |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |