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Autoimmune hepatitis model and medicine screened by using autoimmune hepatitis model

A technology for autoimmunity and hepatitis, applied in the field of non-human animal models, can solve problems such as inability to reflect

Inactive Publication Date: 2018-02-06
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are about 7 models for studying AIH, liver homogenate immunization method, surrogate antigen (Con A / α-GalCer) immunization method, antigen / TCR transgenic method, antigen-loaded DC cell immunization method, virus infection using molecular similarity The method, the specific gene knockout method, and the AIH antigen immunization method are all sensitive models to immunosuppressants, which cannot reflect the 15-20% clinically insensitive patients who are not sensitive to traditional therapies

Method used

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  • Autoimmune hepatitis model and medicine screened by using autoimmune hepatitis model
  • Autoimmune hepatitis model and medicine screened by using autoimmune hepatitis model
  • Autoimmune hepatitis model and medicine screened by using autoimmune hepatitis model

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Experimental program
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Effect test

Embodiment 1

[0037] Tea Tree Mushroom was treated with ddH 2 O soaking, the soaking solution was dialyzed by ammonium sulfate precipitation method to obtain the total protein; then the target protein AAGL was obtained by using the affinity chromatography column of lactose-Sepharose 6B, AAGL 2 Dialyzed overnight in O, the dialysate was freeze-dried, and the SDS-PAGE electrophoresis results of the purified AAGL were as follows figure 1 shown. Dissolve the freeze-dried powder of AAGL in PBS, prepare the concentration of 0.15 mg / ml, 0.2 mg / ml, 0.25 mg / ml, and 0.3 mg / ml, filter it with a 0.22um filter, and inject it into the tail vein according to the weight of the mouse In C57BL / 6 mice, the final injections were 1.5 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 3.0 mg / kg, respectively. Five mice in each group were sacrificed 9 hours later to collect blood, and the serum was separated to measure the contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum;

[0038] Depend on ...

Embodiment 2

[0044] Dissolve the immunosuppressant cyclosporine CsA in olive oil, inject mice intraperitoneally at 50 mg / kg for 24 hours, and then inject mice with 2.5 mg / kg AAGL through the tail vein, 5 mice in each group, and kill the mice 9 hours later. Blood and liver tissue were collected from the rats to detect the changes in ALT and AST levels in the serum; the liver tissue was embedded in paraffin and stained with H&E; at the same time, the Con A model was used as a positive control to detect the efficacy of CsA in inhibiting T cell activation. The specific operation was as follows: After the mice were pretreated with CsA, they were injected with 15 mg / kg Con A through the tail vein, and the mice were sacrificed 8 hours later to collect blood and liver tissue. After the serum was separated, the changes of ALT and AST in the serum were detected; the liver tissue was stained with H&E.

[0045] Depend on image 3 The results of A and B show that the immunosuppressant cyclosporine CsA,...

Embodiment 3

[0049] Download the glycan data of AAGL and Con A version 5.0 from the CFG database, the concentration of the glycan data of AAGL is 200, 100 and 20 μg / ml, the concentration of the data of Con A is 100, 10 and 1 μg / ml; The negative RFU value in the sugar spectrum data is unified as 1, and then the log is calculated 10 (RFU), and finally conduct sugar map heat map analysis according to different sugar chain ends.

[0050] Depend on Figure 4 The results of A show that AAGL can specifically bind sugar chains terminated with galactose, sialylated galactose and sulfonated galactose, while Con A can bind with higher affinity to sugar chains terminated with mannose .

[0051] AAGL and Con A were biotin-labeled with a commercial biotin labeling kit, and BCA was quantified after labeling; mouse spleen lymphocytes were isolated, and the mouse spleen was first separated and ground with rough-edged glass in PBS, and the cell suspension was passed through 200-mesh filter, centrifuge at 3...

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Abstract

The invention discloses an autoimmune hepatitis model and medicine screened by using the autoimmune hepatitis model. Firstly, an AAGL-induced AIH-like mouse hepatitis model is built; the model is notsensitive on the treatment of a conventional immunosuppressor, and can be clinically used as an animal model for conventional immunological suppression treatment on non-sensitive patients. The invention further provides the medicine screened by using the model for treating the autoimmune hepatitis. The result shows that glatiramer acetate can obviously relieve the autoimmune hepatitis symptom of the animal model. The condition indicates that the glatiramer acetate can be used for treating the autoimmune hepatitis. Therefore the invention provides a novel purpose of the glatiramer acetate. Novel medicine is provided for the treatment of the autoimmune hepatitis.

Description

technical field [0001] The invention relates to a non-human animal model, in particular to the establishment of an autoimmune hepatitis model and the use of the drug glatiramer acetate screened by the model in the preparation of drugs for treating autoimmune hepatitis. Background technique [0002] Autoimmune hepatitis (AIH) is an inflammatory disease caused by the disorder of the autoimmune system, causing pathological changes and dysfunction, leading to immune cells directly or indirectly attacking liver parenchymal cells and producing autoantibodies, causing liver damage. The pathological mechanism is still unclear. Current treatment options are limited: immunosuppressive therapy and liver transplantation. Treatment drugs can be divided into standard treatment and elective treatment. The standard treatment is prednisolone combined with or without azathioprine. From 1950 to 1960, corticosteroids and azathioprine were found to have a better effect, but corticosteroids had...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K38/16A61P1/16A61P37/06
CPCA61K38/16A61K49/0008
Inventor 孙慧余文卉
Owner WUHAN UNIV
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