CN107974465B - Slc6a13 gene and application of protein thereof - Google Patents
Slc6a13 gene and application of protein thereof Download PDFInfo
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Abstract
The invention provides an application of an Slc6a13 gene and a protein thereof, and particularly provides a preparation method of a liver injury animal model of a non-human mammal, which comprises the following steps: (1) Providing a cell of a non-human mammal, and inactivating the Slc6a13 gene in the cell to obtain a cell with the Slc6a13 gene inactivated; (2) Preparing a chimeric blastocyst by using the embryonic stem cell with the inactivated Slc6a13 gene and a wild type blastocyst; (3) And preparing a liver injury animal model by using the chimeric blastocyst. The liver injury animal model provided by the invention is an effective acute liver failure animal model, can be used for researching acute liver failure, and can be used for screening and testing specific medicines.
Description
Technical Field
The invention relates to the technical field of biology, in particular to an application of an Slc6a13 gene and a protein thereof.
Background
Acute liver failure refers to the rapid loss of function or death of a large number of hepatocytes from a variety of causes. The disease is a serious clinical syndrome with extremely high mortality. The acute liver failure is caused by various reasons, such as excessive administration of paracetamol, hepatitis virus infection (most commonly hepatitis A virus and hepatitis B virus), excessive drinking, drug idiosyncrasy reaction, anoxic liver injury, autoimmune hepatitis, acute fatty liver of pregnancy, wilson's disease, parasitic infection and the like. In China, the primary cause of liver failure is viral hepatitis (mainly hepatitis B), and the secondary cause is liver injury caused by improper medication and toxic substances. Acute liver failure caused by drugs (such as paracetamol) is the most common in European and American countries.
The current liver failure models are mainly divided into: chemical induction methods including an LPS + galactoside induced acute liver injury model, a CCL4 induced liver injury model, a thioacetamide induced liver injury model, a dimethyl nitrosamine induced liver injury model and the like; induction of drug toxicity: comprises an acetaminophen-induced liver injury model, a tetracycline acetaminophen-induced liver injury model and the like; immune induction methods, including the concanavalin a-induced liver injury model, etc.; the operation method comprises the following steps: such as a liver ischemia reperfusion model, a bile duct ligation-induced liver injury model and the like; and alcohol molding.
Although the application of the current liver injury animal model is wide and the technology is mature, the liver injury animal model is mostly injured due to toxic action, the pathogenesis of the liver injury animal model is different from that of clinical cases, and the target mechanism is unclear.
There is therefore an urgent need in the art to develop an animal model of liver injury in non-human mammals.
Disclosure of Invention
The invention aims to provide a non-human mammal liver injury animal model.
In a first aspect of the invention, there is provided a method of making an animal model of liver injury in a non-human mammal, the method comprising the steps of:
(a) Providing a cell of a non-human mammal, and inactivating the Slc6a13 gene in the cell to obtain a cell with the Slc6a13 gene inactivated;
(b) And (b) preparing the Slc6a13 gene-inactivated liver injury animal model by using the Slc6a13 gene-inactivated cells obtained in the step (a).
In another preferred embodiment, the cells are embryonic stem cells.
In another preferred example, the step (b) further comprises the steps of:
(b1) Preparing a chimeric blastocyst by using the embryonic stem cell with the inactivated Slc6a13 gene and a wild type blastocyst;
(b2) And preparing a liver injury animal model by using the chimeric blastocyst.
In another preferred example, the step (b 2) further comprises the steps of:
(i) Culturing said chimeric blastocyst to develop into a chimeric non-human mammal;
(ii) Mating the chimeric non-human mammal with a wild non-human mammal for breeding, and screening in the offspring to obtain a heterozygote non-human mammal with the Slc6a13 gene inactivated;
(iii) And (3) mutually mating and breeding the heterozygote non-human mammals, and screening the heterozygote non-human mammals with the Slc6a13 gene inactivated in the later generation, namely the liver injury animal model.
In another preferred embodiment, after the step (iii), the method further comprises the steps of:
(iv) Inducing liver damage to said homozygous non-human mammal to obtain said animal model of liver damage.
In another preferred example, the Slc6a13 gene of partial cells in the chimeric blastocyst is inactivated.
In another preferred embodiment, both the heterozygote non-human mammal and the homozygote non-human mammal can reproduce normally.
In another preferred embodiment, the animal model of liver injury has one or more characteristics selected from the group consisting of:
(a) Increase of survival rate
(b) The content of glutamic-pyruvic transaminase or glutamic-oxalacetic transaminase is reduced;
(c) Improvement in liver histopathological symptoms;
(d) Decreased apoptosis of hepatocytes;
(e) In the liver and blood, the expression of inflammatory factors or protein levels is reduced.
In another preferred embodiment, said liver injury inducing conditions are selected from the group consisting of:
LPS + galactoside induction, CCL4 induction, thioacetamide induction, dimethyl nitrosamine induction, acetaminophen induction, tetracycline acetaminophen induction, canavalin A induction, alcohol induction, liver ischemia reperfusion induction, bile duct ligation induction, or a combination thereof.
In another preferred embodiment, the liver injury animal model is selected from the group consisting of: an LPS + galactoside-induced acute liver injury model, a CCL 4-induced liver injury model, a thioacetamide-induced liver injury model, a dimethylnitrosamine-induced liver injury model, an acetaminophen-induced liver injury model, a tetracycline acetaminophen-induced liver injury model, a canavalin a-induced liver injury model, an alcohol-induced liver injury model, a liver ischemia reperfusion model, a bile duct ligation-induced liver injury model, or a combination thereof.
In another preferred embodiment, the non-human mammal is a rodent or primate, preferably including a mouse, rat, rabbit, monkey.
In another preferred embodiment, the liver injury comprises acute liver injury, chronic liver injury, and liver failure.
In another preferred example, said inactivating the Slc6a13 gene comprises gene knockout, gene disruption or gene insertion.
In another preferred example, the gene inactivation comprises that the Slc6a13 gene is not expressed, or that no active Slc6a13 protein is expressed.
In another preferred example, the Slc6a13 gene inactivation comprises liver-specific Slc6a13 gene inactivation or systemic Slc6a13 gene inactivation.
In another preferred example, in step (a), one or more exons in the Slc6a13 gene are deleted or interrupted and replaced with a selection marker using a DNA homologous recombination technique, resulting in a non-human mammalian cell in which the Slc6a13 gene is inactivated.
In another preferred example, exon 2 of the Slc6a13 gene is deleted and replaced with a selection marker.
In another preferred embodiment, the selection marker is selected from the group consisting of: neo gene, TK gene, or a combination thereof.
In a second aspect of the invention, there is provided the use of an animal model prepared by the method of the first aspect of the invention for studying liver damage.
In a third aspect of the invention, there is provided the use of an animal model prepared according to the method of the first aspect of the invention, for preparing an animal model with a high degree of resistance to liver damage.
In a fourth aspect of the invention, there is provided the use of an animal model prepared by the method of the first aspect of the invention for screening or identifying a substance (therapeutic agent) for treating or ameliorating liver damage.
In a fifth aspect of the invention, there is provided a method of screening or identifying potential therapeutic agents for treating or ameliorating liver damage, comprising the steps of:
(a) Administering a test compound to an animal model of liver injury prepared by a method according to the first aspect of the invention in a test group, and determining the severity of liver injury Q1 in said animal model in the test group; administering a control compound (including vehicle) to the animal model of liver injury prepared by the method of the first aspect of the invention in a control group, and measuring the severity of liver injury Q2 in the animal model in the control group;
(b) Comparing the severity Q1 and severity Q2 detected in the previous step to determine whether the test compound is a potential therapeutic agent for treating or ameliorating liver damage;
wherein, if the severity Q1 is significantly lower than the severity Q2, it is indicative that the test compound is a potential therapeutic agent for treating or ameliorating liver injury.
In another preferred embodiment, said reduced severity of liver damage is manifested by: a decrease in the level of glutamate pyruvate transaminase, a decrease in the level of glutamate oxaloacetate transaminase, an improvement in the histopathological symptoms of the liver, a decrease in the apoptotic status of hepatocytes, and/or a decrease in the expression or protein level of inflammatory factors.
In another preferred embodiment, the term "significantly less than" means that the ratio of the severity Q1/severity Q2 is ≦ 1/2, preferably ≦ 1/3, and more preferably ≦ 1/4.
In another preferred embodiment, the method is non-diagnostic and non-therapeutic.
In another preferred embodiment, the test compound and the control compound are administered simultaneously with the induction of liver damage in an animal model of liver damage prepared by the method of the first aspect of the invention.
In a sixth aspect of the invention, there is provided a method of screening for or identifying potential therapeutic agents for treating or ameliorating liver damage, comprising the steps of:
(a) Administering a test compound to a wild-type non-human mammal or cell in a resting state in a test group, detecting the expression of the Slc6a13 gene or protein, or the level of protein transport activity V1; in a control group, a control compound is administered to a wild-type non-human mammal or cell, and the level V2 of expression of the Slc6a13 gene or protein, or protein transport activity, is detected;
(b) Comparing the level V1 and the level V2 detected in the previous step to determine whether the test compound is a potential therapeutic agent for treating or ameliorating liver damage;
wherein, if the level 1 is significantly lower than the level V2, it indicates that the test compound is a potential therapeutic agent for treating or ameliorating liver injury.
In another preferred embodiment, the method is non-diagnostic and non-therapeutic.
In a seventh aspect of the invention, there is provided an animal model of liver damage in a non-human mammal, the model being prepared by a method according to the first aspect of the invention.
In another preferred example, the Slc6a13 gene of the animal model is inactivated.
In an eighth aspect of the invention, there is provided use of an inhibitor of Slc6a13 for the preparation of a formulation or composition for the treatment or alleviation of liver damage.
In another preferred example, the Slc6a13 inhibitor inhibits the expression of the Slc6a13 gene, or inhibits the expression or activity of the Slc6a13 protein.
In another preferred embodiment, the Slc6a13 inhibitor comprises MicroRNA, siRNA, shRNA, or a combination thereof.
In another preferred embodiment, the Slc6a13 inhibitor comprises an antibody.
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.
Drawings
Fig. 1 shows a schematic diagram of the targeting strategy of Slc6a13 knockout mice.
Fig. 2 shows the result of genotyping offspring of the Slc6a13 knockout mouse.
Fig. 3 shows that deletion of the Slc6a13 gene reduced LPS + GalN-induced mouse death.
Fig. 4 shows that the Slc6a13 gene deletion reduces the rise of blood biochemical indicators of liver injury. After the acute liver failure is induced by LPS + GalN for 6 hours, detecting biochemical indexes of blood of the liver injury of the mouse, wherein A: ALT; b: and (5) AST.
FIG. 5 shows that deletion of the Slc6a13 gene ameliorates LPS + GalN-induced pathological impairment of the liver. After the acute liver failure was induced by LPS + GalN for 6H, H & E staining was performed on the liver tissue of each group of mice.
FIG. 5A shows pathological lesions of liver in control WT (left: 40X; right: 400X).
FIG. 5B shows the control group Slc6a13 -/- Pathological lesions of liver (left: 40X; right: 400X).
FIG. 5C shows the hepatic pathological lesions of model group WT (left: 40X; right: 400X).
FIG. 5D shows a model set Slc6a13 -/- Pathological lesions of liver (left: 40X; right: 400X).
Fig. 5E shows the histopathological scoring results.
Detailed Description
The present inventors have extensively and intensively studied to establish an animal model of liver injury, which is a mouse or other non-human mammal in which the Slc6a13 gene is knocked out or inactivated. The animal model is an effective animal model for acute liver failure, can be used for researching the acute liver failure, and can be used for screening and testing specific medicines. The present invention has been completed on the basis of this finding.
Slc6a13 gene and protein thereof
GABA is an important neurotransmitter, and is involved in regulating nerve signal intensity and central nervous system function. GABA receptors and their transporters are also widely distributed in peripheral tissues, suggesting that the gabaergic system has a regulatory role in peripheral tissues. It has now been found that there are 4 GABA transporters in the mammalian genome, named GAT-1, GAT-2, GAT-3, BGT-1 in the human genome and, for the mouse, mGAT1, mGAT3, mGAT4 and mGAT2. The four transporters have some sequence homology with each other, but differ in developmental phase and expression site, suggesting a difference in function. In mice mGAT-1 and mGAT-4 are expressed predominantly in the central nervous system, whereas mGAT-2 and mGAT-3 are expressed predominantly in peripheral tissues. Mouse mGAT-3 corresponds to the human orthologous gene GAT-2, both of which have a homology of up to 91% in protein sequence, and are encoded by a gene named as Slc6a13, and the protein length is 302 amino acid residues. The gene is highly expressed in liver, but no relevant report is available at present on the function of the gene in liver.
Specifically, the Gene ID of the human Slc6a13 Gene is 6540, the Gene ID of the mouse Slc6a13 Gene is 14412, the protein sequence encoded by the human Slc6a13 Gene is shown in SEQ ID NO. 5 (ENSP 00000339260_ Hsap/1-602), and the protein sequence encoded by the mouse Slc6a13 Gene is shown in SEQ ID NO. 6 (ENSMUSP 00000066779_ Mmus/1-602).
In order to analyze the function of the Slc6a13 gene, the invention adopts a gene knockout technology, establishes a Slc6a13 gene knockout mouse, and utilizes the mouse model to carry out phenotypic research, and finds that the deletion of the Slc6a13 gene can relieve acute liver failure induced by bacterial lipopolysaccharide and D-galactosamine (LPS + GalN). The result shows that the Slc6a13 gene deletion mouse can be used as an animal model for researching acute liver injury and acute liver failure, and the coding protein of Slc6a13 can be used as a drug target for screening therapeutic drugs for acute liver injury and acute liver failure. The acute liver failure is clinically severe and is usually caused by viruses and toxicants, the mortality rate is high, and no specific medicine can be used for treating the acute liver failure, so that the research on the disease progression mechanism of the acute liver failure and the development of the specific medicine have important significance.
Inactivation of genes
For the study of a gene of unknown function, many methods are used, such as inactivation of the gene to be studied, analysis of the phenotypic change of the resulting genetic modification, and thus obtaining functional information of the gene. Another advantage of this approach is that it can correlate gene function with disease, thus obtaining both gene function and disease information and animal models of disease that can be treated by the gene as a potential drug or drug target. The method of gene inactivation may be accomplished by means of gene deletion, gene disruption or gene insertion. Among them, gene knockout technology is a very powerful means for studying the functions of human genes as a whole.
Animal model
In the present invention, a very effective non-human mammalian model for studying liver injury is provided.
In the present invention, examples of non-human mammals include (but are not limited to): mouse, rat, rabbit, monkey, etc., more preferably rat and mouse.
As used herein, the term "inactivation of the Slc6a13 gene" includes the case where one or both of the Slc6a13 genes are inactivated, i.e. includes the inactivation of the Slc6a13 gene heterozygously and homozygously. For example, a mouse in which the Slc6a13 gene is inactivated can be a heterozygous or homozygous mouse.
In the present invention, a non-human mammal (e.g., a mouse) in which the Slc6a13 gene is inactivated can be prepared by gene deletion or introduction of a foreign gene (or fragment) to inactivate the Slc6a13 gene. In the art, techniques for inactivating a target gene by gene knockout or introduction of a foreign gene are known, and these conventional techniques can be used in the present invention.
In another preferred embodiment of the invention, the inactivation of the Slc6a13 gene is achieved by gene knockout.
In another preferred embodiment of the invention, the inactivation of the Slc6a13 gene is achieved by inserting a foreign gene (or fragment) into the Slc6a13 gene.
In one embodiment of the invention, a construct containing an exogenous insert can be constructed that contains homology arms homologous to flanking sequences flanking the insertion site of the target gene (Slc 6a 13) such that the exogenous insert (or gene) can be inserted into the Slc6a13 genomic sequence (particularly the exon region) at high frequency by homologous recombination, resulting in a frameshift, premature termination, or knock-out of the mouse Slc6a13 gene, resulting in the deletion or inactivation of Slc6a 13.
The homozygous or heterozygous mouse obtained by the method of the invention can be fertile and normally develop. The inactivated Slc6a13 gene can be inherited to progeny mice on a mendelian basis.
In a preferred embodiment, the invention provides a homozygous mouse model animal lacking the Slc6a13 gene.
Drug candidate or therapeutic agent
In the present invention, there is also provided a method for screening or identifying a candidate drug or therapeutic agent for treating or alleviating liver damage using the animal model of the present invention.
In the present invention, a drug candidate or therapeutic agent refers to a substance known to have a certain pharmacological activity or being tested, which may have a certain pharmacological activity, including but not limited to nucleic acids, proteins, chemically synthesized small or large molecular compounds, cells, and the like. The candidate drug or therapeutic agent may be administered orally, intravenously, intraperitoneally, subcutaneously, or intradermally.
The main advantages of the invention include:
(a) The homozygous or heterozygous animal model obtained by the method can be fertile and normally developed. The transgenic heterozygous male mice have reproductive capacity, and the inactivated Slc6a13 gene can be inherited to offspring mice according to Mendelian rules.
(b) The liver failure animal model can reduce the occurrence of acute liver injury, has high survival rate, and can be used for screening and developing medicaments for treating liver injury, particularly acute liver injury.
(c) According to the invention, a gene knockout method is adopted for the first time to establish an Slc6a13 gene knockout mouse, the expression of the model in acute liver injury is observed, and the result shows that the Slc6a13 gene knockout can obviously reduce the acute liver injury of the mouse caused by the combination of bacterial lipopolysaccharide and D-galactosamine (LPS/GalN).
(d) The invention discloses that the Slc6a13 can be used as a drug action target for the first time.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.
Example 1
Construction of Slc6a13 knockout mice
Construction of mGAT-3 knockout mice A mode of homologous recombination of embryonic stem cell genes of mice was used, and FIG. 1 is a schematic diagram of the gene knockout process. Wherein exon 2 of Slc6a13 in a mouse genome is replaced by a selection gene neo, so that the Slc6a13 gene is damaged, and after the exon is knocked out, a protein initiation codon is knocked out, so that the protein cannot be normally expressed. . The gene targeting is carried out according to the conventional operation, the targeting vector DNA is introduced into a mouse embryonic stem cell (ES) by an electroporation method, cultured on a feeder layer cell consisting of mouse embryonic fibroblasts treated by mitomycin, clone screening is carried out under the action of G418 drugs, PCR identification is carried out on the obtained clones, one PCR primer is respectively designed on the outer side of a homologous recombination arm, the other PCR primer is respectively designed on a screening gene neo gene sequence, PCR products with expected lengths are obtained by PCR on both sides, and the obtained clones are positive clones if the PCR products are correctly verified by sequencing. After cloning and amplifying the positive ES cells, injecting a certain amount of ES cells into a blastocyst cavity of a mouse by using a microinjection method to obtain a chimeric blastocyst, transplanting the chimeric blastocyst into an oviduct of a pseudopregnant mouse to make the pseudopregnant mouse implanted and developed to produce a chimeric mouse, breeding the chimeric mouse and a wild-type mouse to obtain a heterozygote mouse with target gene deletion, and mating the heterozygote mouse to obtain a homozygote mouse with the target gene deletion.
According to the targeting strategy, after the occurrence of correct homologous recombination is verified, the subsequent genotype identification of the mice can adopt a PCR method, and PCR primers are designed as follows:
gGAT3-p1：5’-GAAGGAGGTCACCCTTTTCC-3’；(SEQ ID NO.:1)
gGAT3-p2：5’-AAATCTCCGTCTCCTGCAAA-3’；(SEQ ID NO.:2)
gGAT3-p3：5’-CAACAGATGGCTGGCAACTA-3’；(SEQ ID NO.:3)
gGAT3-p4：5’-TGAATGAACTGCAGGACGAG-3’。(SEQ ID NO.:4)
the results are shown in FIG. 2, where the mice of each genotype used in the experiment were from Slc6a13 +/- And (4) selfing progeny. According to the Slc6a13 gene targeting strategy, primers are designed, and PCR-electrophoresis is carried out on the genome. Different banding results correspond to different genotype mice: the single 400bp band was wild type (+/+); the single 700bp band is a Slc6a13 gene knockout homozygote (-/-); double bands are heterozygotes (+/-).
Example 2
Role of Slc6a13 gene in acute liver injury
Male Wild Type (WT) and Slc6a13 knockout mice (GAT-3) at 10 weeks of age -/- Or slc6a13 -/- ) (homozygous mice) for establishing an acute liver failure model induced by LPS + GalN. All animals need to adapt to the breeding environment for more than one week before the experiment. Mice were injected intraperitoneally with LPS (11. Mu.g/kg body weight) + GalN (700 mg/kg body weight) in combination to induce an acute liver failure model. GalN is a hepatotoxic substance, and its metabolite uridine diphosphate inhibits uracil metabolism, interferes with cellular RNA synthesis, and causes hepatocyte apoptosis. GalN is often used in combination with LPS to induce the acute liver failure model. LPS has strong immunogenicity, and can activate Kupffer cell (a liver-specific macrophage) in liver to attack liver cell and convert GThe damaging effects of alN are further amplified. The model is one of the most common models in the research of acute liver failure at present.
Results show that compared with wild mice, the Slc6a13 knockout mice have obvious resistance to acute liver failure induced by LPS + GalN, and obviously reduced lethality rate (figure 3); blood biochemical markers of alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) were also low (fig. 4).
The results show that Slc6a13 gene knockout can reduce the damage of LPS + GalN to the liver. This inference was verified in pathological examination (FIG. 5), liver H&The E-slice results show: the shape of the liver cells of the control mice is complete and the arrangement is regular (figure 5A, figure 5B); after LPS + GalN administration, WT mice developed massive hepatocyte necrosis and balloon-like degeneration with extensive hepatic sinus congestion (fig. 5C); model group Slc6a13 -/- Mice had less cell necrosis and hyperemia than WT mice (fig. 5D). Histopathological scoring also showed that deletion of the Slc6a13 gene reduced LPS + GalN-induced histopathological damage (p)<0.005 (FIG. 5E).
The results show that the Slc6a13 gene and the protein product thereof play a role in the process of liver injury caused by chemical substances, inhibit the activity of the gene and the protein thereof, and can slow down the occurrence and development of liver injury and liver failure, so that the gene and the protein thereof can be used as drug targets for screening drugs for treating liver injury and liver failure.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes or modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the appended claims of the present application.
Claims (13)
1. A preparation method of a mouse model with acute liver injury is characterized by comprising the following steps:
(a) Providing a mouse cell, and inactivating an Slc6a13 gene in the cell to obtain a cell with an inactivated Slc6a13 gene;
(b) Preparing a mouse model of acute liver injury with the inactivated Slc6a13 gene by using the cells with the inactivated Slc6a13 gene obtained in the step (a),
wherein the protein amino acid sequence coded by the Slc6a13 gene is shown as SEQ ID NO. 6.
2. The method of claim 1, wherein the cell is an embryonic stem cell.
3. The method of claim 1, wherein said step (b) further comprises the steps of:
(b1) Preparing a chimeric blastocyst by using the embryonic stem cell inactivated by the Slc6a13 gene and a wild type blastocyst;
(b2) And preparing a mouse model with acute liver injury by using the chimeric blastocyst.
4. The method of claim 3, wherein said step (b 2) further comprises the steps of:
(i) Culturing the chimeric blastocyst to develop into a chimeric mouse;
(ii) Mating and breeding the chimeric mouse and a wild mouse, and screening in the offspring to obtain a heterozygous mouse with an inactivated Slc6a13 gene;
(iii) And (3) mutually mating and breeding the heterozygote mice, and screening in the later generations to obtain the homozygote mice with the Slc6a13 gene inactivated, namely the acute liver injury mouse model.
5. The method of claim 1, wherein the mouse model of acute liver injury has one or more characteristics compared to a wild-type control mouse selected from the group consisting of:
(a) Survival rate is improved
(b) The content of glutamic-pyruvic transaminase or glutamic-oxalacetic transaminase is reduced;
(c) Improvement in liver histopathological symptoms;
(d) Decreased apoptosis of hepatocytes;
(e) In the liver and blood, the expression of inflammatory factors or protein levels is reduced.
6. The method of claim 5, wherein the acute liver injury inducing condition is LPS + galactoside induction.
7. The method of claim 1, wherein the mouse model of acute liver injury is a LPS + galactoside-induced model of acute liver injury failure.
8. Use of a mouse model prepared by the method of claim 1 to prepare a mouse model with high resistance to acute liver injury.
9. Use of a mouse model prepared by the method of claim 1 to screen for or identify a substance that treats or ameliorates acute liver injury.
10. A method of screening for or identifying a potential therapeutic agent for treating or ameliorating acute liver injury comprising the steps of:
(a) Administering a test compound to a mouse model of acute liver injury prepared by the method of claim 1 in a test group, and detecting the severity of liver injury Q1 in the mouse model in the test group; in a control group, a control compound is applied to the mouse model with acute liver injury prepared by the method of claim 1, and the severity degree Q2 of liver injury of the mouse model in the control group is detected;
(b) Comparing the severity Q1 and severity Q2 detected in the previous step to determine whether the test compound is a potential therapeutic agent for treating or ameliorating acute liver injury;
wherein a severity Q1 that is significantly lower than severity Q2 indicates that the test compound is a potential therapeutic agent for treating or ameliorating acute liver injury.
11. The method of claim 10, wherein the control compound is a vehicle.
12. The method of claim 10, wherein said reduced severity of acute liver injury is manifested by: reduced levels of glutamic-pyruvic transaminase, reduced levels of glutamic-oxaloacetic transaminase, improved histopathological symptoms of the liver, reduced apoptosis of hepatocytes, and/or reduced expression or protein levels of inflammatory factors.
13. The application of the Slc6a13 inhibitor is characterized in that the Slc6a13 inhibitor is used for preparing a preparation for constructing a mouse model of acute liver injury, the Slc6a13 inhibitor inhibits the expression of a Slc6a13 gene or inhibits the expression or activity of a Slc6a13 protein,
wherein the protein amino acid sequence coded by the Slc6a13 gene is shown as SEQ ID NO. 6, and the Slc6a13 protein amino acid sequence is shown as SEQ ID NO. 6.
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