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Uses of Slc6a12 gene and protein thereof

A technology of genes and uses, applied in the biological field, can solve problems such as different pathogenic mechanisms, unclear target mechanisms, and toxic damage

Pending Publication Date: 2018-05-01
SHANGHAI BIOMODEL ORGANISM SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the current animal models of liver injury are widely used and the technology is mature, most of them are injuries caused by toxic effects, the pathogenic mechanism is not the same as that of clinical cases, and the target mechanism is not clear

Method used

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  • Uses of Slc6a12 gene and protein thereof
  • Uses of Slc6a12 gene and protein thereof
  • Uses of Slc6a12 gene and protein thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1 Construction and Identification of Slc6a12 Gene Knockout Mice

[0107] The construction of Slc6a12 knockout mice was carried out according to conventional gene targeting technology. By constructing a targeting vector, it is introduced into embryonic stem cells (ES) (SCR012) extracted from mouse blastocysts, and the Slc6a12 gene is targeted to be destroyed by homologous recombination using the homologous fragment and genome on the targeting vector and the target gene region. figure 1 It is a schematic diagram of the targeting vector and homologous recombination targeting, in which the exon 1-2 region of Slc6a12 in the mouse genome is replaced by the selection gene neo, resulting in the destruction of the Slc6a12 gene and the deletion of the gene start codon.

[0108] The gene targeting experiment was carried out according to routine operations. The targeting vector DNA was introduced into mouse embryonic stem cells (ES) by electroporation, and cultured on feede...

Embodiment 2

[0116] Example 2 The role of Slc6a12 in acute liver injury

[0117] 10-week-old male wild-type (WT) and Slc6a12 knockout mice (slc6a12- / -) were used to establish a model of acute liver failure induced by LPS+GalN. All animals need to adapt to the feeding environment for more than one week before the experiment. Mice were injected intraperitoneally with LPS (11 μg / kg body weight) + GalN (700 mg / kg body weight) to induce an acute liver failure model. GalN is a hepatotoxic substance, and its metabolite uridine diphosphate can inhibit uracil metabolism, interfere with cellular RNA synthesis, and lead to hepatocyte apoptosis. GalN is often combined with LPS to induce acute liver failure models. LPS has strong immunogenicity and can activate Kupffer cells in the liver (a liver-specific macrophage) to attack liver cells, further expanding the damage of GalN. This model is currently one of the most commonly used models in the study of acute liver failure.

[0118] For the survival...

Embodiment 3

[0122] Embodiment 3 uses the drug verification drug screening platform for the treatment of liver injury (such as acute liver injury)

[0123] In this example, the model animal mice established in Example 1 were injected with tiopronin and / or bicyclol, which are current clinical drugs for treating acute liver injury, and then the severity of liver injury in the model animal mice was evaluated.

[0124] The result shows that medicine tiopronin and / or bicyclol etc. can significantly reduce the liver damage severity of model animal mice, can significantly improve the survival rate of model animal mice, significantly reduce the blood index of liver damage (such as alanine aminotransferase and Aspartate aminotransferase content), significantly improve the liver histopathological symptoms, significantly reduce the apoptosis of liver cells, and significantly reduce the expression levels of inflammatory factors or their proteins in the liver and blood.

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Abstract

The present invention provides uses of a Slc6a12 gene and protein thereof, specifically a preparation method of a non-human mammal liver injury animal model. The preparation method comprises: (a) providing non-human mammal cells, and inactivating the Slc6a12 gene in the cells to obtain Slc6a12 gene-inactivated cells; and (b) preparing a Slc6a12 gene-inactivated liver injury animal model by using the Slc6a12 gene-inactivated cells obtained in the step (a). According to the present invention, the obtained non-human mammal liver injury animal model is an effective acute liver injury animal model,can be used for studying acute liver injury, and can be used for screening and testing of specific drugs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to the use of the Slc6a12 gene and its protein. Background technique [0002] Acute liver failure refers to the rapid loss of function or death of a large number of liver cells caused by various reasons. The disease is a severe clinical syndrome with a very high mortality rate. There are many causes of acute liver failure, such as overdose of paracetamol, hepatitis virus infection (most common in hepatitis A and B), excessive drinking, idiosyncratic drug reactions, hypoxic liver injury, autoimmune hepatitis, pregnancy Acute fatty liver, Wilson's disease, parasitic infection, etc. In my country, the primary cause of liver failure is viral hepatitis (mainly hepatitis B), followed by liver damage caused by improper medication and toxic substances. In European and American countries, acute liver failure caused by drugs (such as paracetamol) is the most common. [...

Claims

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Application Information

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IPC IPC(8): C12N15/877A01K67/027C12Q1/6883G01N33/68G01N33/50
CPCA01K67/0271G01N33/5008G01N33/6893C12N15/877C12Q1/6883C07K14/47C12Q2600/158A01K2267/0393A01K2207/12A01K2227/10A01K2217/075G01N2800/085
Inventor 刘震泽孙瑞林匡颖
Owner SHANGHAI BIOMODEL ORGANISM SCI & TECH DEV
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