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Construction method and application of non-alcoholic steatohepatitis mouse model based on PEDF/LDLR double-gene knockout

A gene knockout mouse, steatohepatitis technology, applied in the field of medicine, can solve the problems of lack of NASH models, etc., and achieve the effect of wide promotion and simple method

Pending Publication Date: 2022-05-31
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no recognized NASH model that can reflect the comorbidities of cardiovascular diseases in NASH patients

Method used

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  • Construction method and application of non-alcoholic steatohepatitis mouse model based on PEDF/LDLR double-gene knockout
  • Construction method and application of non-alcoholic steatohepatitis mouse model based on PEDF/LDLR double-gene knockout
  • Construction method and application of non-alcoholic steatohepatitis mouse model based on PEDF/LDLR double-gene knockout

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] In this example, the body weight, blood lipid content, glucose tolerance, insulin tolerance, serum insulin content and blood sugar of the above-mentioned four groups of animals were measured.

[0033] The above 4 groups of animals were weighed at 0 weeks, 16 weeks, 32 weeks and 48 weeks respectively, and fasted for 12-16 hours before weighing; glucose tolerance test and insulin tolerance test were performed at 32 weeks of induction. The specific operations were as follows: Fasting for 12-16 hours, 1 g / kg glucose or 1 U / kg insulin were injected intraperitoneally, and blood glucose was measured at 0, 15, 30, 60, 90, 120, and 180 minutes after injection, and the blood glucose change curve was drawn. Calculate the area under the curve; at 32 weeks and 48 of induction, the animals were fasted for 12-16 hours, blood glucose was measured by a blood glucose meter, and the mice were dissected to collect serum samples, and commercial kits were used to detect serum total cholestero...

Embodiment 2

[0037] In this example, the liver weight, liver-to-body ratio, liver function enzyme level, liver steatosis, liver inflammatory response and liver fibrosis of the above-mentioned four groups of animals were evaluated at 32 weeks of induction.

[0038] At the 32nd week of induction, animals were fasted for 12-16 hours and then sacrificed by anesthesia. Serum and tissue samples were collected, and liver tissue was weighed; serum alanine aminotransferase and aspartate aminotransferase levels were detected using commercial kits; Sections were stained with HE and Masson to evaluate liver lesions.

[0039] The result is as figure 2 As shown, where A: gross picture of liver; B: liver weight; C: liver body ratio; D: serum alanine aminotransferase level; E: serum aspartate aminotransferase level; F: HE staining; G: liver steatosis score; H: Liver ballooning degeneration score; I: liver lobular inflammation score; J: Masson staining; K: liver fibrosis score; *, p<0.05, vs wild-type no...

Embodiment 3

[0045] In this example, the liver weight, liver-to-body ratio, liver function enzyme level, liver steatosis, liver inflammatory response and liver fibrosis of the above-mentioned four groups of animals were evaluated at 48 weeks of induction.

[0046] At 48 weeks of induction, animals were fasted for 12-16 hours and then sacrificed by anesthesia, serum and tissue samples were collected, and liver tissue was weighed; serum alanine aminotransferase and aspartate aminotransferase levels were detected using commercial kits; Sections were stained with HE and Masson to evaluate liver lesions.

[0047] The result is as image 3 As shown, where A: liver weight; B: liver body ratio; C: serum alanine aminotransferase level; D: serum aspartate aminotransferase level; E: HE staining; F: CD45 immunohistochemical staining; G: liver steatosis score; H: liver ballooning degeneration score; I: liver lobular inflammation score; J: Masson staining; K: liver fibrosis score; *, p<0.05, vs wild-ty...

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Abstract

The invention discloses a construction method and application of a non-alcoholic steatohepatitis mouse model based on PEDF / LDLR double-gene knockout, and belongs to the technical field of medicines. High fat diet is adopted to induce PEDF / LDLR double-gene knockout mice to generate the phenotype of non-alcoholic steatohepatitis, and the constructed animal model shows the characteristics of obesity, hyperlipidemia, insulin resistance, hyperglycemia, liver fatty degeneration, liver inflammatory response, liver fibrosis, liver tumor, cardiovascular injury and the like; the method can be used for preclinical research on research and development of drugs for metabolic diseases such as non-alcoholic steatohepatitis and the like. The construction method of the animal model is simple and easy to implement and can be widely popularized. The model can better simulate the morbidity characteristics, the lesion process and the lesion characteristics of all stages of human NASH, and can reflect the coexisting characteristics of cardiovascular diseases of NASH patients, so that an important tool is provided for preclinical research of research and development of drugs for metabolic diseases such as NASH and the like.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for constructing a mouse model of nonalcoholic steatohepatitis based on PEDF / LDLR double gene knockout and its application. Background technique [0002] Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by steatosis and lipid storage in liver cells without a history of excessive alcohol consumption. It has become the main cause of chronic liver disease worldwide. The pathological changes include simple fatty liver, non-alcoholic steatohepatitis (NASH), liver fibrosis, and eventually develop into liver cirrhosis and hepatocellular carcinoma with the progression of the disease. NASH is an extreme form of nonalcoholic fatty liver development, defined as the appearance of steatosis accompanied by inflammation and liver cell damage, and is characterized by liver steatosis, inflammation, liver cell damage, and varying degree...

Claims

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Application Information

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IPC IPC(8): A01K67/027
CPCA01K67/0276A01K2207/25A01K2217/077A01K2227/105A01K2267/035
Inventor 蔡卫斌李兴会吴燕笛
Owner SUN YAT SEN UNIV
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