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Construction method of gene immune induced Graves' Ophthalmopathy animal model and application of rapamycin type medicine

A technology of immune induction and animal model, which is applied in the field of biomedicine, can solve problems such as deficiencies, and achieve the effect of simple operation and good animal reactivity

Active Publication Date: 2020-06-12
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, the research on GD animal models has made important progress, among which the TSHR-adenovirus model is widely respected in terms of success rate and replicability; GO animal models are also being explored and tried continuously, including TSHR-plasmid-assisted Protocols of electroporation and TSHR-adenovirus multiple injections, but insufficient in reproducing hyperthyroid state and ocular disease state, respectively

Method used

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  • Construction method of gene immune induced Graves' Ophthalmopathy animal model and application of rapamycin type medicine
  • Construction method of gene immune induced Graves' Ophthalmopathy animal model and application of rapamycin type medicine
  • Construction method of gene immune induced Graves' Ophthalmopathy animal model and application of rapamycin type medicine

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Experimental program
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Effect test

experiment example 1

[0056] Experimental case 1. Preparation of animal model of thyroid-related eye disease

[0057] Experimental equipment: Ad-TSHR (purchased from Shenzhen Baienwei Biotechnology Co., Ltd.), PBS, female 6-8 weeks old inbred BALB / C mice (purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.)

[0058] Preparation:

[0059] 1. The adenovirus (Ad-TSHR) stock solution expressing human thyroid stimulating hormone receptor is diluted with PBS according to the volume ratio of 1:50;

[0060] 2. 6-week-old healthy female BALB / C mice were randomly divided into two groups, the control group (8 mice) and the modeling group (22 mice).

[0061] 3. Mice in the control group were injected with adenovirus expressing green fluorescent protein (Ad-EGFP); mice in the model group were injected with adenovirus with extracellular fragment of TSH receptor (Ad-TSHR). The dose of each injection was 10^8 pfu, and 25ul was injected into the left and right gluteus maximus muscles respe...

experiment example 2

[0075] Experimental case 2. Evaluation of therapeutic effect of rapamycin on animal model of thyroid-related ophthalmopathy

[0076] Experimental equipment: Ad-TSHR (purchased from Shenzhen Baienwei Biotechnology Co., Ltd.), PBS, female 6-8 weeks old inbred BALB / C mice (purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.), Rapamycin (purchased from Shanghai Dibo Biotechnology Co., Ltd.)

[0077] experimental method:

[0078]1. Add rapamycin powder to mouse feed in the form of enteric-coated particles (14ppm);

[0079] 2. The 6-week-old healthy female BALB / C mice were randomly divided into three groups: normal control group, common model group, and rapamycin group;

[0080] 3. The common model group and the rapamycin group were stimulated by the animal model of thyroid-associated eye disease induced by gene immunity, and the normal control group was stimulated by injection of adenovirus (Ad-EGFP) expressing green fluorescent protein. The normal contro...

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Abstract

The invention discloses a construction method of a gene immune induced Graves' Ophthalmopathy animal model and application of a rapamycin type medicine. The construction method comprises the followingsteps: 1) diluting a mother liquid of an adenovirus expressing a thyroid-stimulating hormone receptor by using PBS (phosphate buffer saline) according to a volume ratio of 1:50 so as to obtain an Ad-TSHR (adenovirus-thyroid-stimulating hormone receptor) diluted liquid; 2) on the first day of a first week, respectively injecting 25[mu] L of the Ad-TSHR diluted liquid into left gluteus maximus muscle and right gluteus maximus muscle of experiment animals; and 3) repeating operation of the step 2) at the 3rd, 10th, 14th, 18th, 22nd, 26th and 30th weeks till the experiment animals are induced tohave Graves' Ophthalmopathy, so as to successfully construct the gene immune induced Graves' Ophthalmopathy animal model. The construction method makes up the blank of the field of Graves' Ophthalmopathy models, and is a novel exploration on rapamycin in treating Graves' Ophthalmopathy.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a method for constructing an animal model of thyroid-related eye disease induced by gene immunity and the application of rapamycin drugs. Background technique [0002] Graves' disease (GD), also known as toxic diffuse goiter, is an organ-specific autoimmune disease whose pathogenesis is that the body produces thyroid-stimulating hormone receptor (TSHR) thyrotropin receptor antibody (TRAb). [0003] Thyroid-related ophthalmopathy, also known as Graves' Ophthalmopathy (GO), is a common extrathyroidal complication of Graves' disease, with an incidence of 20%-50% in patients with Graves' disease. As an autoimmune-mediated inflammatory disease, GO occurs when the body breaks the immune tolerance of T cells to self-antigens under the combined action of genetics and the environment, and then antigen-specific T cells develop specificity in the thyroid and orbital tissues. Cross-react...

Claims

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Application Information

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IPC IPC(8): A01K67/027A61K31/436A61P27/02
CPCA01K67/027A61K31/436A61P27/02A01K2267/0387
Inventor 王悦施秉银张萌伍丽萍
Owner THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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