Method of establishing gnotobiotic animal obesity model and application of model

An obese model, a technology of living animals, which is applied in the field of constructing obesity models of living animals, and can solve problems such as not very clear, unclear mechanism, etc.

Active Publication Date: 2012-11-28
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As we all know, obesity has become an increasingly serious public health problem in modern society. However, so far, the mechanism by which gut flora participates in the occurrence and development of metabolic diseases such as obesity and insulin resistance is not very clear. Specifically, gut flora Is it the cause or the result of metabolic diseases, what kind of bacteria can cause metabolic diseases, and what kind of bacteria can improve obesity, insulin resistance and other symptoms is not very clear

Method used

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  • Method of establishing gnotobiotic animal obesity model and application of model
  • Method of establishing gnotobiotic animal obesity model and application of model
  • Method of establishing gnotobiotic animal obesity model and application of model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation and identification of Enterobacter cloacae B29 strain

[0030] Analysis of the intestinal flora of obese people found that the structure of intestinal flora in most severely obese patients is seriously unbalanced. For example, an extremely obese volunteer (26 years old, male, Han nationality, weight 174.9kg, BMI 58.78kg / m2) The proportion of Enterobacter in the intestinal flora structure of) is as high as 35% (16S rRNA gene clone library analysis). At the same time, the physical examination data also showed that the volunteer had severe metabolic syndrome. The volunteer underwent a 23-week nutritional intervention (composition: whole grains, foods from the same source of medicine and food, and prebiotics). After 9 weeks, the volunteer's weight dropped by 30.1kg, and after 23 weeks, it dropped by 51.4kg. The symptoms of the disease have also improved. The analysis of the intestinal flora showed that the proportion of Enterobacter in the intestinal flor...

Embodiment 2

[0038] Example 2 Strain B29 was stably colonized in the intestinal tract of sterile mice.

[0039] During the experiment, fresh stool samples were collected every two weeks, immediately weighed accurately, and diluted 10-fold with sterile 0.01M phosphate (PBS) buffer. The formula of 0.01M phosphate (PBS) buffer is as follows: 135mM sodium chloride (NaCl), 2.7mM potassium chloride (KCl), 1.5mM potassium dihydrogen phosphate (KH 2 PO 4 ), and 8mM dipotassium hydrogen phosphate (K 2 HPO 4 ), pH 7.2. Spread 100 μl of the suspension on an LB plate, culture at 37°C for 16 hours, and count the colonies. The results showed that the density of Enterobacter cloacae B29 strain in the fecal samples of model mice (ie high-fat diet+B29) and normal diet+B29 mice during the experiment period (0-16 weeks) could reach 10 10 -10 12 / g wet weight stool sample, showing that Enterobacter cloacae B29 strain can be stably colonized in the intestinal tract of sterile mice (see figure 1 ).

[0040] Based o...

Embodiment 3 29

[0041] Example 3 B29 strain causes obesity and insulin resistance in sterile mice with high fat diet.

[0042] During the experiment, the body weight of each mouse was measured once a week and accurately weighed with an electronic balance (d=0.01). In the 16th week of the experiment, the mice were given oral glucose (2g / kg body weight) after fasting for 5 hours, and blood was taken from the tail-tip vein before taking glucose (0 minutes) and at 15, 30, 60 and 120 minutes after taking glucose. , Immediately use the Roche blood glucose meter to measure the blood glucose concentration for oral glucose tolerance test. After taking glucose for 120 minutes, collect a 50μl blood sample from the eye socket, leave it at room temperature for 1 hour, 3000 revolutions / separate the heart for 15 minutes, collect the serum, use an ELISA kit (Mercodia, Uppsala, Sweden) to detect the blood insulin content, 2 hours after the meal Determination of insulin levels. After 16 weeks of the experiment,...

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Abstract

The invention provides a method of establishing a gnotobiotic animal obesity model and an application of the model. According to the method, a gnotobiotic animal obesity model is established by inoculating an LPS (lipopolysaccharides) producing strain under a high-fat diet condition. When a germfree animal is fed with high-fat diet, a microorganism such as LPS producing strain or a combination thereof is inoculated, then the microorganism can colonize in the intestinal tract of the germfree animal, and the animal shows symptoms of disorder of the systems of metabolism, immunity and the like such as obesity, insulin resistance and chronic inflammation at the same time. The invention also relates to a method for screening the microorganism involved in obesity and combination thereof and providing a drug target for treating obesity and a method for screening the microorganism with an effect of preventing and treating metabolic diseases such as obesity, and provides a new method for treating obesity; and the method provided by the invention can be applied to the screening and effect evaluation on foods (formula), medicines, nourishments, healthcare products, beverages and the like with an effect of preventing and treating metabolic diseases such as obesity.

Description

Technical field [0001] The present invention relates to biotechnology, in particular to a method for constructing an obesity model in animals that have been born, and its use in screening of microorganisms involved in the occurrence of obesity, screening of microorganisms that prevent and treat obesity, as well as foods, formulas, drugs, nutrients, health products, and beverages In the application. Background technique [0002] There are a large number of symbiotic microorganisms living in the human body. Its status and function are equivalent to an important "organ" acquired after birth, which plays an irreplaceable role in human health. As a complete human body, eukaryotic cells account for only 10% of the total number of cells. , And 90% are prokaryotic cells. Therefore, Nobel Laureate Lederberg proposed that the human body is a "super organism" composed of eukaryotic cells and prokaryotic cells symbiotically in the body. There are more than 1,000 types of symbiotic microorgan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02A61K49/00
CPCA01K67/02A61K49/00A61K49/0008A01K2207/12A01K2267/0368A01K2267/0337A01K67/0271A01K2207/25A01K2227/105A01K2267/0362A01K67/027
Inventor 赵立平费娜
Owner SHANGHAI JIAO TONG UNIV
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