CN108498522A - A kind of fast modeling method of young rat type 1 diabetes - Google Patents
A kind of fast modeling method of young rat type 1 diabetes Download PDFInfo
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- CN108498522A CN108498522A CN201810650602.XA CN201810650602A CN108498522A CN 108498522 A CN108498522 A CN 108498522A CN 201810650602 A CN201810650602 A CN 201810650602A CN 108498522 A CN108498522 A CN 108498522A
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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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7008—Compounds having an amino group directly attached to a carbon atom of the saccharide radical, e.g. D-galactosamine, ranimustine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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Abstract
The invention discloses a kind of fast modeling methods for the young rat type 1 diabetes for belonging to biotechnology.The method, including steps are as follows：To the young mice tail vein injection streptozotocin after fasting, taboo water, injection volume is 50 60mgkg‑1Once, injection 1 time daily, continuous injection 4 days, model is established in induction.The present invention provides a kind of effectively and rapidly method for the modeling of young mice type 1 diabetes.The method of the present invention uses tail vein injection, and in conjunction with specific STZ dosage, operability is strong, convenient and practical, can succeed and quickly establish the young rat T1DM models with typical clinical symptom.
Description
Technical field
The invention belongs to biotechnologies, and in particular to a kind of fast modeling method of young rat type 1 diabetes.
Background technology
Type 1 diabetes, original name insulin-dependent diabetes mellitus mostly occur in Children and teenager, can also betide various
Age.Onset compares drastically, and internal insulin is absolutely insufficient, is easy to happen ketoacidosis, it is necessary to use insulin therapy ability
Satisfactory effect is obtained, otherwise by threat to life.Type 1 diabetes are the autoimmune diseases for threatening human health, are studied at present
The result shows that type 1 diabetes are mainly by T cell mediateds, attack destroys beta Cell of islet, leads to hypoinsulinism, from
And cause blood glucose rise.
Current 98% Children Diabetes are type 1 diabetes, and the annual morbidity in China is 1.04/10 ten thousand.In recent years epidemic disease
It learns studies have shown that the incidence of global type 1 diabetes increases year by year, the 4-6 Sui and 10-14 Sui Gao Fa Nian Ling for type 1 diabetes.
Childhood be critical period that body is in physiology and psychological development, and the unstable condition of Children Diabetes is easy to fluctuate, need
Lifelong dietary control and insulin injection are wanted, this brings various spirit worried and financial burden to patient and family.
There is researcher to be showed using the delayed type 1 diabetes (T1DM) of multiple low dose streptozotocin (STZ) induction
Mice Body endolymph cell can be activated for STZ, inflammation of pancreatic islet sample pathological change similar with mankind T1DM is generated in 2-3 weeks, and
The pancreas islet pathology damage prompt STZ of inflammatory may be by the diabetes of cellular immune pathway induction, to establish and the mankind
The similar mouse T1DM models of T1DM height.People have carried out greatly the mechanism and therapy of type 1 diabetes in recent years
Experimental study is measured, but model is confined to adult rats, young rat type 1 diabetes model modeling success rate is low, and young rat is easy death,
The foundation of young rat type 1 diabetes model is of great significance to children T1DM clinical researches and treatment.
Invention content
It is an object of the invention to overcome the problems of the prior art, a kind of fast run-up of young rat type 1 diabetes is provided
Mould method.
For this purpose, the technical scheme is that：
A kind of fast modeling method of young rat type 1 diabetes, including steps are as follows：Prohibit to young mice before modeling
Food prohibits water, by the young mice tail vein injection streptozotocin after fasting, taboo water, injection volume 50-60mgkg-1Once,
Injection 1 time daily, continuous injection 4 days, model is established in induction.
In above-mentioned fast modeling method, it is preferred that the injection volume is 50-55mgkg-1Once.
In above-mentioned fast modeling method, most preferably, the injection volume is 50mgkg-1Once.
In above-mentioned fast modeling method, dosage needed for young mice is calculated by empty body weight.
In above-mentioned fast modeling method, the fasting, the method for prohibiting water are：To young mice fasting, taboo water before modeling
12-18h。
In above-mentioned fast modeling method, the method further includes：Adaptable fed young mice 3-4 days before modeling.
In above-mentioned fast modeling method, the young mice is the mouse less than 3 week old.
In above-mentioned fast modeling method, the young mice is the mouse of 3 week old.
In above-mentioned fast modeling method, the mouse is C57BL/6 mouse.
In above-mentioned fast modeling method, the 12nd day blood sugar monitoring carried out for three days on end since modeling, blood sugar concentration >=
16.7mmol·L-1Person is to model successfully.
Beneficial effects of the present invention are：
The present invention provides a kind of effectively and rapidly method for the modeling of young mice type 1 diabetes.The method of the present invention is adopted
With tail vein injection, in conjunction with specific STZ dosage, operability is strong, convenient and practical, can succeed and quickly establish and face with typical case
The young rat T1DM models of bed symptom.Such method can be with low dosage, few induction modeling, and modeling process mouse death rate
Low, high at mould rate, mouse has typical diabetic symptom after Cheng Mo.
Description of the drawings
Fig. 1 is the type 1 diabetes model mice outside drawing of various dose STZ inductions, A 30mgkg-1Group, B are
50mg·kg-1Group.
Fig. 2 is the weight and change of blood sugar curve graph of the type 1 diabetes model mice of various dose STZ inductions.
Fig. 3 is the Pancreas pathology figure of the type 1 diabetes model mice of various dose STZ inductions：A is 30mgkg-1Group
Pancreas HE colored graphs, B 50mgkg-1The pancreas HE colored graphs of group, C 30mgkg-1Group is insulin immunohistochemical visualization
Figure, D 50mgkg-1Group insulin immunohistochemical visualization figure.
Specific implementation mode
With reference to specific embodiment, the present invention will be further described, but the scope of protection of present invention is not limited to
In the range of embodiment statement, anyone can obtain other various forms of products under the inspiration of the present invention, however,
Make any variation in its shape or component ratio, it is every that there is technical solution identical or similar to the present application, all fall within this
In the protection domain of invention.Material used in the present invention and device are unless otherwise specified commercially available.
1 modeling method of embodiment
By the C57BL/6 mouse of 3 week old after adaptable fed 3d, overbit nail label.Modeling starts preceding fasting, prohibits water
15h weighs empty body weight and blood glucose and records.It is randomly divided into blank control group, 30mgkg-1Group, 40mgkg-1Group,
50mg·kg-1Group, 60mgkg-1Every group 10, dosage needed for mouse is calculated by empty body weight for group.
STZ is dissolved in pH4.2-4.5 sodium citrate buffer solutions (in biology work clean bench 4 DEG C after 0.22 μm of membrane filtration
Seal up for safekeeping) in, prepare the solution of a concentration of 500 ㎎/ml.Abundant dissolved drug solution is kept in dark place, and is positioned on ice,
Use finishes in 30min, now with the current every time, invalid to prevent drug degradation.Medication is：By mouse tail vein injection.The
After shot STZ, free diet is recovered immediately；Bearing Mice Life sign and existing state are observed at any time.It is 1 time/d, continuous to inject
4d。
2 model of embodiment sets up judgment criteria
To the continuous 3d surveys random blood sugar of mouse (i.e. start since modeling to the 12nd day) after modeling 1w, continuous 3d is equal >=
16.7mmol·L-1Person is to model successfully, and measure each group mouse blood sugar and weight weekly.
The results show that observation mouse general signs, there is the glycosuria of more foods, more drinks, diuresis " more than three " in each experimental mice
Sick classical symptom, chaeta smoothness decline, and skin is slack and undisciplined, and build is become thin, and anthropometic delays；Occur apathetic or hyperfunction
Equal nervous system abnormalities performance, as shown in FIG. 1, FIG. 1 is 30mgkg for profile variation-1Group and 50mgkg-1Mouse in group
Outline drawing.
Wherein 50mgkg-1、60mg·kg-1The continuous 3d of mouse blood sugar of experimental group >=16.7mmolL-1, it is modeled as
Work(, wherein 50mgkg-1 experimental mices diabetic symptom are most typical as shown in Fig. 2, 50mgkg-1Group blood glucose is obviously high
In 30mgkg-1Group, 30mgkg-1Group blood glucose is not up at mould standard, as shown in Fig. 2, 40mgkg-1Group effect and
30mg·kg-1Group effect is not much different.70mg·kg-1Group mouse is all dead early period in modeling, influences later stage scale-model investigation.
3 HE of embodiment dyeing and Immunohistochemical Method detection Pancreas pathology change
Target organ Pathologic specimen is collected after modeling 1 month.Cervical dislocation puts to death mouse, wins pancreas, kidney rapidly,
It is fixed in 4% paraformaldehyde solution, routine paraffin wax embedding, slice.Islet tissue is divided into two parts, and a part carries out HE dyes
Color, microscopically observation islet tissue pathological change.Another part tissue does Immunohistochemical detection, and islet tissue is cut
Piece carries out the dewaxing of dimethylbenzene gradient, Gradient elution using ethanol, distillation water washing, and antigen retrieval is incubated antibody and DAB colour developings, bush
Element is redyed, and transparent, neutral gum mounting is dehydrated.It micro- sem observation and takes pictures.
By 30mgkg-1Group and 50mgkg-1Group mouse presses above-mentioned experiment process, and as a result display is as shown in figure 3, pancreas islet
HE dyes visible 50mgkg-1Group islet area is compared with 30mgkg-1Group substantially reduces, and the enhancing of cell cytosol eosinophilic staining goes out
Existing karyopycnosis, as shown in Figure 3.
Insulin Showed by immune group result, 30mgkg-1Group islet tissue is full, and cell distribution is uniform, pancreas islet uniformly dyeing
Color is positive brown color, color depth；And 50mgkg-1Group pancreas islet volume is obviously reduced, insulin secretion two groups of reductions earlier above, such as
Shown in Fig. 3.
Result above illustrates that few secondary, low dosage STZ is successfully established mouse T1DM, and such method by tail vein injection
The model of induction has typical diabetic symptom, is convenient for experimental study.
Claims (10)
1. a kind of fast modeling method of young rat type 1 diabetes, which is characterized in that including steps are as follows：To children before modeling
Water is prohibited in year mouse fasting, by the young mice tail vein injection streptozotocin after fasting, taboo water, injection volume 50-60mg
kg-1Once, injection 1 time daily, continuous injection 4 days, model is established in induction.
2. fast modeling method according to claim 1, which is characterized in that the injection volume is 50-55mgkg-1One
It is secondary.
3. fast modeling method according to claim 2, which is characterized in that the injection volume is 50mgkg-1Once.
4. according to claim 1-3 any one of them fast modeling methods, which is characterized in that it is small to calculate childhood by empty body weight
Dosage needed for mouse.
5. according to claim 1-3 any one of them fast modeling methods, which is characterized in that the fasting, the method for prohibiting water
For：To young mice fasting, taboo water 12-18h before modeling.
6. according to claim 1-3 any one of them fast modeling methods, which is characterized in that the method further includes：It is building
Adaptable fed young mice 3-4 days before mould.
7. according to claim 1-6 any one of them fast modeling methods, which is characterized in that the young mice is less than 3
The mouse of week old.
8. fast modeling method according to claim 7, which is characterized in that the young mice is the mouse of 3 week old.
9. fast modeling method according to claim 7 or 8, which is characterized in that the mouse is C57BL/6 mouse.
10. fast modeling method according to claim 1, which is characterized in that carry out for three days on end within the 12nd day since modeling
Blood sugar monitoring, blood sugar concentration >=16.7mmolL-1Person is to model successfully.
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| CN201810650602.XA CN108498522A (en) | 2018-06-22 | 2018-06-22 | A kind of fast modeling method of young rat type 1 diabetes |
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| CN201810650602.XA CN108498522A (en) | 2018-06-22 | 2018-06-22 | A kind of fast modeling method of young rat type 1 diabetes |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111066728A (en) * | 2020-01-02 | 2020-04-28 | 康美华大基因技术有限公司 | Construction method of hyperglycemia animal model |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104116724A (en) * | 2014-06-24 | 2014-10-29 | 上海交通大学医学院附属瑞金医院 | Application of FFAR2 natural ligand acetate in preparation of medicine for treating type 1 diabetes |
| CN106377529A (en) * | 2016-11-10 | 2017-02-08 | 广西中医药大学 | Method for quickly making models of diabetic mice |
-
2018
- 2018-06-22 CN CN201810650602.XA patent/CN108498522A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104116724A (en) * | 2014-06-24 | 2014-10-29 | 上海交通大学医学院附属瑞金医院 | Application of FFAR2 natural ligand acetate in preparation of medicine for treating type 1 diabetes |
| CN106377529A (en) * | 2016-11-10 | 2017-02-08 | 广西中医药大学 | Method for quickly making models of diabetic mice |
Non-Patent Citations (2)
| Title |
|---|
| 宋旸 等: "STZ诱导C57BL小鼠T1DM模型优化的研究", 《中国实验诊断学》 * |
| 李甜 等: "链脲佐菌素诱导C57小鼠1型糖尿病模型的研究", 《现代生物医学进展》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111066728A (en) * | 2020-01-02 | 2020-04-28 | 康美华大基因技术有限公司 | Construction method of hyperglycemia animal model |
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Application publication date: 20180907 |
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