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Construction method of low-quality articular cartilage animal model based on epigenetic modification and application of construction method

A technology of epigenetic modification and animal model, which is applied in the field of construction of low-quality articular cartilage animal models, can solve problems such as multi-generational genetic animal models of chondrodysplasia, and achieve the effect of simple and easy method

Pending Publication Date: 2020-03-31
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of epidemiological investigations and our previous experimental studies have shown that adult osteoarthritis has a fetal origin. However, there is no report on a multigenerational genetic animal model of chondrodysplasia caused by adverse environmental exposure during pregnancy.

Method used

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  • Construction method of low-quality articular cartilage animal model based on epigenetic modification and application of construction method
  • Construction method of low-quality articular cartilage animal model based on epigenetic modification and application of construction method
  • Construction method of low-quality articular cartilage animal model based on epigenetic modification and application of construction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] [Example 1] Construction of a low-quality articular cartilage animal model based on epigenetic modification

[0056] 1. Construction method

[0057]SPF grade normal Wistar rats were purchased from Hubei Provincial Center for Disease Control and Prevention, animal license number: SCXK (Hubei) 2012-2014. This study was approved by the Ethics Committee of Wuhan University Medical Center, and was carried out in strict accordance with the relevant treatment guidelines of international laboratory animal protection certification and evaluation institutions. The experimental animals were kept in a barrier environment with a temperature of 22-25° C. and a humidity of 50%, with 12 hours of day and night alternation. 12-week-old female Wistar rats (180-220g) and males (260-300g) were caged together at a ratio of 2:1 for males and females every night. On the next day, the vaginal secretion smears of the female rats were checked, and the vaginal plug observed under the microscope ...

Embodiment 2

[0073] [Example 2] Using the low-quality articular cartilage animal model based on epigenetic modification constructed in Example 1 of the present invention to explore the epigenetic mechanism (applied to the study of the mechanism of fetal-derived adult osteoarthritis)

[0074] 1. Method steps

[0075] 1.1 According to the method in Example 1, a low-quality articular cartilage animal model based on epigenetic modification was established.

[0076] 1.2 Use the RT-qPCR method to detect the TGFβ signaling pathway indicators of F1 female offspring in utero and F1 / F2 / F3 female offspring 12 weeks after birth, including TGFβ, TGFβR1, SMAD2, SMAD3, and SOX9. RT-qPCR experimental steps and primers used refer to Example 1.

[0077] 1.3 Use immunohistochemical methods to detect the TGFβ signaling pathway indicators in F1 female offspring and F1 / F2 / F3 female offspring 12 weeks after birth, including TGFβ, TGFβR1, SMAD2, and SOX9. Refer to Example 1 for immunohistochemical methods and e...

Embodiment 3

[0085] [Example 3] Application of the low-quality articular cartilage animal model based on epigenetic modification constructed in Example 1 of the present invention in the papain damage experiment

[0086] 1. Method steps

[0087] 1.1 According to the method in Example 1, a low-quality articular cartilage animal model based on epigenetic modification was established.

[0088] 1.2 F3 progeny of 12-week-old female rats, two litters of rats were taken from the control group and the dexamethasone-treated group (PDE group).

[0089] 1.3 Papain damage test: Inject 4% papain and normal saline solution into the left knee joint cavity of each rat, each injection volume is 0.1mL, inject every other day (1.3.5.7), treat for 1 week, and observe for 2 weeks; After 15 weeks, the knee joints of the rats were taken, stained with Safranin O Fast Green, and the cartilage quality of the knee joints was evaluated using the Mankin's scoring system. The Mankin's scoring system consists of four e...

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Abstract

The invention provides a construction method of a low-quality articular cartilage animal model based on epigenetic modification and an application of the construction method. The method comprises thefollowing steps: giving 0.1-2.0 mg / kg dexamethasone per day to perform subcutaneous treatment during the 9-20 days of pregnancy through the normal pregnant rats, enabling the pregnant rats to freely eat food and perform natural production to obtain F1 filial generation, and further obtaining F2, F3,..., Fn filial generation; and obtaining a low-quality articular cartilage animal model based on epigenetic modification according to the detected related indexes of the knee joint cartilage quality of the F1, F2, F3 and Fn filial generations. The construction method provided by the invention is simple, high in success rate, effective, reliable and high in repeatability, reduces the influence caused by manual operation factors, can be used for exploring an epigenetic mechanism, articular cartilage damage factors and osteoarthritis prevention and treatment drug screening, and has important clinical significance.

Description

technical field [0001] The invention belongs to the technical field of animal model construction, and in particular relates to a construction method and application of a low-quality articular cartilage animal model based on epigenetic modification. Background technique [0002] Osteoarthritis (osteoarthritis) is one of the common senile degenerative diseases. Synovial lesions at the morphological and biochemical levels. In recent years, with the aging of the society and the impact of bad living habits, the incidence of osteoarthritis has been increasing year by year, and it has become one of the main sports system diseases that affect people's quality of life, bringing a heavy burden to patients, their families and society . However, the etiology of osteoarthritis is still unclear, and most of the drugs used clinically are only for pain relief, but not for cure. Therefore, the pathogenesis of osteoarthritis and its therapeutic drugs have become the focus of scientific res...

Claims

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Application Information

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IPC IPC(8): G16B5/00G16B20/40G16B25/00G16B40/00
CPCG16B5/00G16B20/40G16B25/00G16B40/00
Inventor 陈廖斌汪晖谭杨铁楷李庆贤
Owner WUHAN UNIV
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