CN102812921A - Method for establishing type 2 diabetes animal model and application of type 2 diabetes animal model in screening of blood sugar reducing medicaments - Google Patents
Method for establishing type 2 diabetes animal model and application of type 2 diabetes animal model in screening of blood sugar reducing medicaments Download PDFInfo
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- CN102812921A CN102812921A CN2012103218095A CN201210321809A CN102812921A CN 102812921 A CN102812921 A CN 102812921A CN 2012103218095 A CN2012103218095 A CN 2012103218095A CN 201210321809 A CN201210321809 A CN 201210321809A CN 102812921 A CN102812921 A CN 102812921A
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Abstract
The invention provides a method for establishing a type 2 diabetes animal model. An animal is fed with a high-fat and high-sugar feed for a period of time, and the animal is drenched with alcohol with certain concentration at the same time. The attack process of human type 2 diabetes is simulated in the method; and the prepared animal model has typical characteristics of the type 2 diabetes such as high fasting blood glucose, high insulinemia, hyperlipidemia and impaired glucose tolerance, and is suitable for mechanism research of the type 2 diabetes and screening of a large quantity of medicaments.
Description
Technical field
The present invention relates to a kind of method for preparing the diabetes B animal model and the application in the diabetes medicament screening thereof, be specifically related to a kind ofly be used for analyzing the pathogenetic animal model preparation method of diabetes B and in the application of the animal model that carries out the screening of diabetes B related drugs.
Background technology
Diabetes; The traditional Chinese medical science is referred to as diabete; Be that what to cause owing to the absolute or relative hyposecretion of insulin in the body is a kind of endocrine system disease of master with metabolism disorders such as sugar, fat, protein; Can involve each organ of whole body and tissue, cause diabetic complication, be one of chronic disease of serious harm society and human health.
The onset diabetes rate is in rising trend in recent years, and according to World Health Organization's statistics, the onset diabetes number reached 1.35 hundred million in 1998, reached 1.6 hundred million in 2000, reached 2.4 hundred million by 2004.Show according to nearest epidemiology survey, the onset diabetes rate of China by 0.67% before 15 years rise in recent years 3.2%, wherein the diabetes B patient of non-insulin-depending type accounts for more than 90% of sum.
Existing hypoglycemic research comprises molecular level, cellular level, live body level; Wherein maximum with the viviperception meaning.Living Animal Models mainly comprises: the spontaneous type diabetes animal model, such as the ob/ob mouse; The diabetes animal model that chemicals causes, such as mouse or rat diabetes model that Streptozotocin causes, these animal models all are the characteristics of simulating human diabetes.In addition, also have the high sugar associating of high fat chemicals to set up diabetes animal model.
There are some researches show that type-II diabetes and diet have important relatedly with drinking, and can not reflect diet and alcohol to the pathogenic influence of diabetes B in the above-mentioned model.
Once there was report to do the high sugar of too high fat and united construction diabetes B animal model with alcohol; Because there is defective in the high sugar prescription of its high fat; Lack sodium taurocholate (promoting muroid animal cholesterol and fat absorption), cause part index number not statistically significant in the animal model, modeling is success not.
Summary of the invention
The purpose of this invention is to provide a kind of method that makes up the diabetes B animal model; The pathogenic process of this method simulating human diabetes B; The characteristic feature of diabetes Bs such as high fasting blood-glucose, hyperinsulinemia, high fat of blood, sugar tolerance attenuating appears in prepared animal model, is suitable for diabetes B excitation research and carries out a large amount of drug screening.
The inventive method is characterised in that high-sugar-fat-diet nutrition purposes, especially for feeding animals a period of time, irritates clothes finite concentration alcohol to animal simultaneously, and animal blood glucose rising, blood fat are raise, and insulinemia and insulin resistance take place.
The inventive method may further comprise the steps:
Through freely ingesting, continue to feed animal pattern 2-10 week with high-sugar-fat-diet, irritate the edible alcohol 1-20 ml/kg that the body of stomach integration is counted 10%-60% simultaneously.
In the inventive method, high-sugar-fat-diet is made up of basal feed, sucrose, grease, cholesterol, sodium taurocholate, yolk powder.Wherein, grease can be lard, perhaps butter.
The ratio of high-sugar-fat-diet is basal feed 50%-70%; Sucrose 10%-20%; Grease 10%-20%; Cholesterol 1%-5%; Sodium taurocholate 0.5%-2%; Yolk powder 5%-10%.
Animal pattern comprises wild-type mice, like the C57BL/6J mouse, and BALB/c mouse, ICR mouse, KM mouse; The wild type rat, like the Wistar rat, SD rat etc.
Animal pattern is after feeding for 4 to 20 weeks, and daytime, promptly morning, 6:00 began jejunitasly not cut off the water supply on an empty stomach, and 18:00 put to death and got blood evening; Automatic biochemistry analyzer detects each item index in the blood, like blood sugar, and insulin, blood fat; Cholesterol, low-density lipoprotein, HDL.
Description of drawings
Fig. 1 is a rat OGTT result of the test, ● expression experimental group data, ▲ expression normal group experimental data, * representes and the normal group contrast that P < 0.05.
Embodiment
1, mouse test:
C57BL mouse, is divided into two groups, each 15 at random by 30.Normal group is fed normal feed, ad lib drinking-water; Experimental group is fed high-sugar-fat-diet, freely drinks water, and irritates stomach 0.2 ml 30% edible alcohol every day.
The ratio of high-sugar-fat-diet is a basal feed 60%; Sucrose 15%; Grease 15%; Cholesterol 3%; Sodium taurocholate 0.5%; Yolk powder 6.5%.
After raising for 8 weeks, daytime on an empty stomach, promptly morning 6:00 on an empty stomach, evening, 18 broken ends were got blood, separation of serum, automatic biochemistry analyzer detects each item index in the blood.Statistics software analysis data significance.
The result is as shown in table 1; With the normal group contrast, the fasting blood-glucose of experimental group mouse, insulin, cholesterol, triglycerides, low-density lipoprotein significantly increase, and HDL significantly reduces; Consistent with human diabetes B patient's physiochemical indice trend, the modeling success.
Table 1 mouse modeling result
| Group | Normal group | Experimental group |
| Fasting blood-glucose (mM) | 4.25±0.35 | 14.23±3.9 |
| Insulin (pM) | 186.54 ± 10.76 | 268.85 ± 12.01 △ |
| Cholesterol (mM) | 2.49±0.21 | 3.36±0.33 |
| Triglycerides (mM) | 1.18 ± 0.17 | 1.92±0.32 |
| HDL (mM) | 1.69 ± 0.12 | 2.26±0.14 |
| Low-density lipoprotein (mM) | 3.26 ± 0.27 | 1.5 ± 0.41 |
Data are expressed as: mean+SD; With compared with normal,
△Expression P<0.05.
2, rat test:
The SD rat, 30 are divided into two groups at random, each 15.Normal group is fed normal feed, ad lib drinking-water; Experimental group is fed high-sugar-fat-diet, freely drinks water, and irritates stomach 1 ml 35% edible alcohol every day.
The ratio of high-sugar-fat-diet is a basal feed 60%; Sucrose 15%; Grease 15%; Cholesterol 2%; Sodium taurocholate 1.5%; Yolk powder 6.5%.
After raising for 8 weeks, on 12 hours on an empty stomach daytime, broken end is got blood, separation of serum, and automatic biochemistry analyzer detects each item index in the blood.
The result is as shown in table 2; With the normal group contrast, the fasting blood-glucose of experimental group mouse, insulin, cholesterol, triglycerides, low-density lipoprotein significantly increase, and HDL significantly reduces; Consistent with human diabetes B patient's physiochemical indice trend, the modeling success.
Table 2 rat modeling result
| Group | Normal group | Experimental group |
| FPG (mM) | 4.5±0.32 | 15.2±1.12 △ |
| Fins (pM) | 216.54 ± 18.17 | 288.51 ± 31.11 △ |
| TC (mM) | 1.27 ± 0.25 | 2.75 ± 0.61 △ |
| TG (mM) | 0.98±0.08 | 1.81±0.29 △ |
| HDL（mM） | 1.26±0.32 | 0.78 ± 0.16 △ |
| LDL（mM） | 1.06±0.10 | 2.98±0.23 △ |
Data are expressed as: mean+SD; With compared with normal,
△Expression P<0.05.
3, rat OGTT test
The SD rat, 30 are divided into two groups at random, each 15.Normal group is fed normal feed, ad lib drinking-water; Experimental group is fed high-sugar-fat-diet, freely drinks water, and irritates 1 milliliter of 35% edible alcohol of stomach every day.
The ratio of high-sugar-fat-diet is a basal feed 60%; Sucrose 15%; Grease 15%; Cholesterol 2%; Sodium taurocholate 1.5%; Yolk powder 6.5%.
After raising for 8 weeks, irritate stomach 2g/kg glucose 12 hours on an empty stomach daytime, use blood glucose meter to measure rat blood sugar respectively at 0 minute, 30 minutes, 60 minutes, 90 minutes, 120 minutes.The result is shown in accompanying drawing 1, and (▲) compares with normal group, and experimental group (●) sugar tolerance significantly reduces.With the normal group contrast, the oral glucose tolerance of rats in test groups significantly reduces, and blood sugar significantly increased in each time period, the modeling success.
Claims (4)
1. the preparation method of a diabetes B animal model and the application in the screening hypoglycemic drug thereof is characterized in that feeding high-sugar-fat-diet, irritate the stomach edible alcohol simultaneously; Obtain the diabetes B animal model, wherein the prescription of high-sugar-fat-diet is basal feed 50%-70%, sucrose 10%-20%; Grease 10%-20%; Cholesterol 1%-5%, sodium taurocholate 0.5%-2%, yolk powder 5%-10%; The edible alcohol concentration of irritating stomach is volume fraction 10% to 40%, and irritating body of stomach long-pending is 0.1 milliliter to 2 milliliters.
2. the preparation method of diabetes B animal model according to claim 1 is characterized in that the letting animals feed cycle was 4 to 20 weeks.
3. the preparation method of diabetes B animal model according to claim 1 is characterized in that the animal of using comprises C57BL mouse, BALB/c mouse, ICR mouse, KM mouse, Wistar rat, SD rat.
4. the preparation method of diabetes B animal model according to claim 1; It is characterized in that animal pattern after feeding a period of time, get 12 hours on an empty stomach daytime before the blood, promptly morning, 6:00 began jejunitasly not cut off the water supply; 18:00 put to death and got blood evening, detected the relevant physiochemical indice of diabetes.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103314925A (en) * | 2013-06-21 | 2013-09-25 | 四川大学华西医院 | Method for preparing rhesus monkey type 2 diabetic model |
| CN103461660A (en) * | 2013-09-17 | 2013-12-25 | 上海中医药大学附属曙光医院 | Diabetes-inducing high-fat feed and application thereof to preparation of diabetic foot ulcer rat experimental model |
| CN103463135A (en) * | 2013-09-17 | 2013-12-25 | 上海中医药大学附属曙光医院 | Method and reagent kit for establishing experimental model of diabetic foot ulcer rat with mixed infection of Gram positive bacteria and negative bacteria |
| CN104886339A (en) * | 2015-06-23 | 2015-09-09 | 南通特洛菲饲料科技有限公司 | High-fat high-glucose model feed |
| CN106135133A (en) * | 2016-08-02 | 2016-11-23 | 四川大学 | The construction method of checking glucose resultant index animal experimental model and the animal model of structure thereof |
| CN107182937A (en) * | 2017-07-13 | 2017-09-22 | 广州市中医医院 | A kind of construction method of internal oxygen-starved prediabetes animal model |
| CN108619162A (en) * | 2017-03-24 | 2018-10-09 | 贵州医科大学 | The construction method of type 1 diabetes mouse responsible drinking model |
| CN111557273A (en) * | 2020-07-02 | 2020-08-21 | 贵州中医药大学 | Method for inducing type 2 diabetes animal model by low temperature and diet rhythm regulation |
| CN112997928A (en) * | 2021-02-22 | 2021-06-22 | 温州大学 | Construction method and application of type II diabetic zebra fish model |
| CN115024279A (en) * | 2022-07-26 | 2022-09-09 | 陆辉强 | Construction method of zebra fish diabetic vasculopathy model |
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2012
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103314925A (en) * | 2013-06-21 | 2013-09-25 | 四川大学华西医院 | Method for preparing rhesus monkey type 2 diabetic model |
| WO2014202022A1 (en) * | 2013-06-21 | 2014-12-24 | 四川大学华西医院 | Method for preparing rhesus macaque diabetes mellitus type 2 model |
| CN103461660A (en) * | 2013-09-17 | 2013-12-25 | 上海中医药大学附属曙光医院 | Diabetes-inducing high-fat feed and application thereof to preparation of diabetic foot ulcer rat experimental model |
| CN103463135A (en) * | 2013-09-17 | 2013-12-25 | 上海中医药大学附属曙光医院 | Method and reagent kit for establishing experimental model of diabetic foot ulcer rat with mixed infection of Gram positive bacteria and negative bacteria |
| CN103461660B (en) * | 2013-09-17 | 2014-12-31 | 上海中医药大学附属曙光医院 | Diabetes-inducing high-fat feed and application thereof to preparation of diabetic foot ulcer rat experimental model |
| CN104886339A (en) * | 2015-06-23 | 2015-09-09 | 南通特洛菲饲料科技有限公司 | High-fat high-glucose model feed |
| CN106135133A (en) * | 2016-08-02 | 2016-11-23 | 四川大学 | The construction method of checking glucose resultant index animal experimental model and the animal model of structure thereof |
| CN108619162A (en) * | 2017-03-24 | 2018-10-09 | 贵州医科大学 | The construction method of type 1 diabetes mouse responsible drinking model |
| CN107182937A (en) * | 2017-07-13 | 2017-09-22 | 广州市中医医院 | A kind of construction method of internal oxygen-starved prediabetes animal model |
| CN107182937B (en) * | 2017-07-13 | 2021-07-16 | 广州市中医医院 | Construction method of in-vivo hypoxia-type diabetes mellitus early-stage animal model |
| CN111557273A (en) * | 2020-07-02 | 2020-08-21 | 贵州中医药大学 | Method for inducing type 2 diabetes animal model by low temperature and diet rhythm regulation |
| CN111557273B (en) * | 2020-07-02 | 2022-02-08 | 贵州中医药大学 | Method for inducing type 2 diabetes animal model by low temperature and diet rhythm regulation and application thereof in diabetes treatment |
| CN112997928A (en) * | 2021-02-22 | 2021-06-22 | 温州大学 | Construction method and application of type II diabetic zebra fish model |
| CN115024279A (en) * | 2022-07-26 | 2022-09-09 | 陆辉强 | Construction method of zebra fish diabetic vasculopathy model |
| CN115024279B (en) * | 2022-07-26 | 2024-04-12 | 陆辉强 | Construction method of zebra fish diabetic vasculopathy model |
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