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A method for constructing an animal model of thyroid-associated eye disease induced by gene immunity and application of rapamycin

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: 2022-03-25
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
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  • Application Information

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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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  • A method for constructing an animal model of thyroid-associated eye disease induced by gene immunity and application of rapamycin
  • A method for constructing an animal model of thyroid-associated eye disease induced by gene immunity and application of rapamycin
  • A method for constructing an animal model of thyroid-associated eye disease induced by gene immunity and application of rapamycin

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Experimental program
Comparison scheme
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 microparticles (14ppm);

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

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

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Abstract

The invention discloses a method for constructing an animal model of thyroid-associated eye disease induced by gene immunity and application of rapamycin drugs. The construction method is as follows: 1) Dilute the stock solution of adenovirus expressing human thyroid-stimulating hormone receptor with PBS at a volume ratio of 1:50 to obtain Ad-TSHR dilution; 2) On the first day of the 0th week, give the experimental The left and right gluteus maximus of the animal were injected with 25uL Ad-TSHR dilution respectively; 3) the operation of step 2) was repeated in the 3rd, 6th, 10th, 14th, 18th, 22nd, 26th, and 30th weeks until the experimental animals were induced to produce Thyroid-associated eye disease, that is, the successful construction of an animal model of thyroid-associated eye disease induced by gene immunity. The invention fills up the gap in the field of thyroid-associated eye disease models, and is also a new exploration of rapamycin for the treatment of thyroid-associated eye diseases.

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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/33
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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