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Expression method of autophagy gene MAP1LC3 in rabbit hormonal femoral head avascular necrosis

A technology for femoral head necrosis and autophagy genes, applied in the fields of biochemical equipment and methods, microbiological determination/inspection, material inspection products, etc.

Pending Publication Date: 2019-12-13
内蒙古医科大学第二附属医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there have been no published research reports on the role of autophagic programmed cell death in SANFH

Method used

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  • Expression method of autophagy gene MAP1LC3 in rabbit hormonal femoral head avascular necrosis
  • Expression method of autophagy gene MAP1LC3 in rabbit hormonal femoral head avascular necrosis
  • Expression method of autophagy gene MAP1LC3 in rabbit hormonal femoral head avascular necrosis

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Experimental program
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Effect test

experiment example

[0100] Experimental example: 1. Modeling and methods

[0101] 1.1 Experimental materials

[0102] 1.1.1 Experimental animal health 36 5-month-old New Zealand big-eared white rabbits, body weight (2.0-2.5) kg, male or female, standard diet, reared in a single cage. Purchased from Dilepu Biological Resources Development Co., Ltd., Xi’an, Shaanxi Province, certificate number: 61001600000083

[0103] 1.1.2 Experimental Instruments

[0104]

[0105] Other equipment used in the experiment was provided by the Key Laboratory of Molecular Pathology, Inner Mongolia Medical University.

[0106] 1.1.3 Main reagents

[0107]

[0108] Routine reagents such as HE staining, ethylenediaminetetraacetic acid decalcification solution, and absolute ethanol were provided by the Key Laboratory of Molecular Pathology, Inner Mongolia Medical University.

[0109] 1.1.4 Preparation of main reagents

[0110] 1) Preparation of PBS buffer:

[0111]

[0112]

[0113] Dissolve in 1000mL ult...

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Abstract

The invention relates to an expression method of an autophagy gene MAP1LC3 in rabbit hormonal femoral head ischemic necrosis, which comprises the following steps: inducing a hormonal femoral head ischemic necrosis animal model by using hormone, and detecting the influence of hormone on the mRNA and protein high expression of osteocytes and osteoblasts MAP1LC3; discussing 3-methyladenine; regulating and controlling the mRNA and protein high expression of osteocytes, osteoblast autophagic death and osteocytes and osteoblast MAP1LC3, and showing that when mRNA of the MAP1LC3 in a group B is highly expressed, the expression quantity of mRNA of the MAP1LC3 in a group C is lower than that of mRNA of the MAP1LC3 in the group B in the first, second, third and fourth periods; and showing that whenMAP1LC3 protein in the group B is highly expressed in the first and the second periods, MAP1LC3 protein expression in the group C is obviously reduced. Immunofluorescence staining is characterized inthat the GFP-LC3 fusion protein expression of osteocytes and osteoblasts in the group B is higher than that of a control group, and the GFP-LC3 fusion protein expression of the group C is obviously reduced. Wherein the hormone causes autophagic cell death of osteocytes and osteoblasts in a SANFH model; and high expression of mRNA and protein of MAP1LC3 of osteocytes and osteoblasts is caused. The3-MA inhibits death of osteocytes and osteoblast autophagic cells.

Description

technical field [0001] The invention relates to the research field of steroid-induced avascular necrosis of femoral head, in particular to an expression method of autophagy gene MAP1LC3 in steroid-induced avascular necrosis of femoral head in rabbits. Background technique [0002] Steroid-induced Avascular Necrosis of Femoral Head (SANFH) is a difficult problem for the medical community in China and even the world. There are many reasons for the avascular necrosis of the femoral head. The avascular necrosis of the femoral head caused by the use or short-term use of large doses of corticosteroids (hereinafter referred to as hormones) has puzzled the entire medical community. The biological process of hormone-induced avascular necrosis of the femoral head is quite complicated, and its pathogenesis has not been clearly defined until today, and it is even more helpless for its ideal treatment clinically. Beginning in 1957, the pathogenesis of avascular necrosis of the femoral h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883G01N33/6893C12Q2600/158G01N2800/10
Inventor 赵振群刘万林白锐龚瑜林赵爱青王文选孟晨阳
Owner 内蒙古医科大学第二附属医院
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