Preparation method for novel non-alcoholic fatty liver disease model

A non-alcoholic and fatty liver disease technology, applied in the field of human non-alcoholic fatty liver disease (NAFLD) model preparation, can solve the problems of large developmental differences, difficult to control, high sugar, etc., to achieve strong pertinence, promote occurrence, and reasonable route Effect

Active Publication Date: 2013-12-25
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The advantage is to explore the mechanism of a certain factor in the pathogenesis of human NAFLD; the disadvantage is high sugar, high fat or gene knockout, which is quite different from the occurrence and development of human NAFLD, and it is difficult to control
At present, there is no literature showing the establishment of a non-gene knockout, non-high-fat, non-high-glucose metabolic regulation model of NAFLD, and the establishment of this model is more beneficial for exploring the mechanism of abnormal fat metabolism and promoting the prevention and treatment of NAFLD

Method used

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  • Preparation method for novel non-alcoholic fatty liver disease model
  • Preparation method for novel non-alcoholic fatty liver disease model
  • Preparation method for novel non-alcoholic fatty liver disease model

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The livers of the model mice and control mice were compared after 2 weeks, and the size and shape of the livers were as follows: Figure 5 Shown: Fatty liver animal model sees massive hepatic fat accumulation ( Figure 5 A). There was no change in the rat liver of the control group ( Figure 5 B). Visually, there was a clear difference in liver size.

Embodiment 2

[0052] After the two groups of mice were treated, 5ml of blood was collected from the hearts of the mice, and the serum was separated in time and stored at -20°C; the liver was removed from the mice, and the liver tissue was divided into 1. 5 cm × 1. 5 cm × 0. 3 cm , respectively placed in AAF mixed fixative solution (ethanol, acetic acid, neutral formaldehyde, V / V / V, refer to the method introduced by Xu Ping (Journal of Henan Medical University, 1999, 34 (4): 75) for fixation.

[0053] The liver tissues of the two groups of mice were routinely frozen into sections using a Leica-CM1900 cryostat and Feather disposable blades. The samples were collected, embedded in ordinary glue (frozen), sectioned, and sealed after staining.

[0054] Routine pathological examination (H&E staining) was performed on the mice of the two groups; the liver tissue of the mice was stained with oil red O method for fat accumulation, and the fat accumulation in the liver tissue was dynamically analyzed....

Embodiment 3

[0066] Dynamic analysis of liver tissue fat. The liver tissue of the two groups of rats after staining was dynamically analyzed, and the obtained dynamic analysis of liver tissue fat was compared, and it was obtained that during the establishment of the dynamic change model of fatty acids and total fatty acids, fat accumulation in liver tissue, long-chain fatty acids, short-chain fatty acids and Total fatty acid level (attached table); changes in peripheral blood, long-chain fatty acid, short-chain fatty acid and total fatty acid levels. For example, the changes of fatty acid concentration in mouse liver tissue are as follows:

[0067] The change of fatty acid concentration in rat liver in the attached table for two weeks (n=5, nMol / gram wet weight liver tissue)

[0068] group total fatty acids free fatty acid short chain fatty acids long chain fatty acids model mouse 392.16±53.08* 252.28±28.21* 61.56±23.28* 78.32±9.51* Control mice 323.78±5...

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Abstract

The invention discloses a preparation method for a novel non-alcoholic fatty liver disease model. The preparation method comprises the following steps: a, randomly dividing fed mice into two groups, with one group being a model group and the other group being control mice, recording states everyday and taking urine, hepatic tissue and blood for lipid analysis; b, processing experimental subjects; c, building a model; d, collecting hepatic tissue and blood; e, preparing conventional frozen sections from hepatic tissue specimens of the mice, taking materials, wrapping the frozen sections with common glue and then successively carrying out dyeing and sealing; f, carrying out conventional pathological examination on model mice; g, carrying out conventional pathological examination on control mice; and h, comparing results of dynamic analysis of hepatic tissue fat obtained in step f and step g. The novel non-alcoholic fatty liver disease model established in the invention has the advantages of reasonable route, easy operation and strong pertinency and is an effective tool for further research on the non-alcoholic fatty liver disease.

Description

technical field [0001] The invention relates to a method for preparing a human non-alcoholic fatty liver disease (NAFLD) model. Specifically, it can be used for analysis of the mechanism of human NAFLD, evaluation of diagnostic methods, drug screening and prevention, and belongs to the medical field. Background technique [0002] Non-alcoholic fatty liver disease (NAFLD) is divided into two categories: primary and secondary. The former is related to insulin resistance and genetic susceptibility, while the latter is caused by some special reasons. Excessive weight gain and overweight caused by overnutrition, obesity, diabetes, hyperlipidemia and other metabolic syndrome-related fatty liver, and cryptogenic fatty liver all belong to the category of primary non-alcoholic fatty liver disease; while nutritional Fatty liver caused by malnutrition, total parenteral nutrition, rapid weight loss after bariatric surgery, drug, environmental and industrial poisoning belongs to the cat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/205
Inventor 姚登福
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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