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Modeling method of Sjogren syndrome mouse model

A technology of Sjögren's syndrome and mouse model, applied in the field of medical experiments, can solve the problems of low molding rate, difficult research, long cycle, etc., and achieve the effects of short modeling time, easy death and high molding rate.

Inactive Publication Date: 2016-11-16
魏伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many organ damages in spontaneous animal models are quite similar to those of Sjögren's syndrome in humans, the high cost, long cycle, difficult control and low modeling rate make laboratory research encounter many difficulties
At present, there are deficiencies in several induced animal models at home and abroad. For example, the use of rats to create models requires high economic costs, and the long cycle will cause the functions of various tissues and organs of rats to gradually decline, thereby affecting the accuracy of the experiment. sex
In addition, the route, dose, time, frequency and selection of adjuvant induced by mouse antigen will all affect the success rate of the model.

Method used

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  • Modeling method of Sjogren syndrome mouse model

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Embodiment

[0028] 1 Preparation of submandibular gland antigen:

[0029] Take 4 C57BL / 6 mice, kill them by dislodging their cervical vertebrae, place them in bromogeramine for disinfection for 15 minutes, and aseptically remove the bilateral submandibular glands in an ultra-clean bench, separate the capsule and connective tissue, wash them with PBS, and put them into a sterile penicillin vial Add 2ml of PBS to the bottle, cut the submandibular gland in the bottle with ophthalmic scissors, fully homogenize it with an electric homogenizer in an ice bath, transfer it to an EP tube and centrifuge at 3000r / 15min / 4°C, discard The upper layer of lipid was removed, and the supernatant was taken for BCA protein quantification. The antigen concentration was adjusted to 4 mg / ml with PBS, and an equal amount of Freund's complete adjuvant (FCA) or Freund's incomplete adjuvant (FIA) was added. Dilute the antigen concentration to 2mg / ml, and pipette repeatedly until the two liquids dissolve into each o...

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Abstract

The invention discloses a modeling method of an Sjogren syndrome mouse model. The modeling method comprises the following steps: killing mice, taking out bilateral salivary glands and peeling off capsules and connective tissues; washing with PBS (Poly Butylenes Succinate); adding the PBS according to the amount of adding 0.5ml of the PBS into each salivary gland; shearing the salivary glands into pieces, and uniformly homogenizing and centrifuging in an ice bath; then taking supernatant and quantifying salivary gland antigens by adopting a BCA (Bicinchoninic Acid) protein quantifying method; adjusting the concentration of the salivary gland antigens to be 4mg / ml by utilizing the PBS; adding equal quantity of an FCA (Freund Complete Adjuvant) or an FIA (Freund Incomplete Adjuvant) and diluting the concentration to be 2mg / ml; repeatedly blowing and beating until two liquid phases are dissolved mutually to form an ivory color; randomly grouping C57BL / 6 mice and shaving off furs on the backs of the mice; carrying out intradermal multi-site injection of 2mg / ml mouse salivary gland antigens prepared by the FCA on the back and tails of the mice on the current day and the 7th day, wherein the injection amount is 0.1ml per mouse; injecting equal quantity of the salivary gland antigens prepared by the FIA on the 14th day through the same method; after modeling for about 6 weeks, detecting indexes and screening the successfully modeled mice. The modeling method disclosed by the invention is high in modeling efficiency and short in modeling time.

Description

technical field [0001] The invention relates to the field of medical experiments, in particular to a method for making a mouse model of Sjogren's syndrome. Background technique [0002] Sjögren syndrome (SS) is a chronic inflammatory autoimmune disease characterized by invasion of salivary and lacrimal glands. Sjögren's syndrome can be divided into primary and secondary according to the cause of the disease. Primary Sjögren's syndrome (primary Sjogren Syndrome, pSS) is a global disease that occurs more frequently in women and the elderly, with a male to female ratio of about 1:9 to 1:20. Dry mouth and dry eyes are the typical clinical symptoms of the disease, and some patients also have intermittent parotitis and later rampant dental caries; in addition, it includes other atypical extraglandular system symptoms. [0003] At present, the main animal models of Sjögren's syndrome at home and abroad are mice, including two types, namely spontaneous and induced. Spontaneous an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00
CPCA61K39/0008A61K2039/55566
Inventor 魏伟吴华勋顾芳
Owner 魏伟
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