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Construction method of animal model of rat with compressive injury of nerve roots

A construction method and animal model technology, applied in the field of medical animal model construction, can solve problems such as no animal models, and achieve the effects of high success rate, unique construction method, and simple technical operation

Inactive Publication Date: 2021-01-15
厦门市中医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the prior art, there is no animal model that can meet the above research requirements

Method used

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  • Construction method of animal model of rat with compressive injury of nerve roots
  • Construction method of animal model of rat with compressive injury of nerve roots
  • Construction method of animal model of rat with compressive injury of nerve roots

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 120 male SD rats (2 months old) were randomly divided into four groups: normal group (normal), sham operation group (shamsurgery), nerve root exposed group (nerve root exposed) and nerve root compressed group (nerve root compressed), in

[0035] In the nerve root compression group, the rats were anesthetized by intraperitoneal injection of ketamine, and a 4 cm midline incision was made with a scalpel as a reference. The left paraspinal muscles were dissected and contracted to expose the left lamina of L5. Part of the left lamina and articular process of L5 was gradually dissected to expose the intact nerve root. will be like figure 1 A rubber block shown in A (a right-angled triangle, the lengths of the two right-angled sides are 4 μm and 2 μm respectively, and the thickness is 1 μm) is inserted into the spinal canal and pressed on the L5 nerve root (such as figure 1 shown in B). When the rubber block can no longer be inserted, the incision is stitched layer by laye...

Embodiment 2

[0041] The DRG obtained in Example 1 was fixed with 4% paraformaldehyde for 8 hours, dehydrated with gradient sucrose, embedded in OCT gel, sectioned at 6 μm, stained with hematoxylin and eosin, and then analyzed with the imaging analysis system with CMIAS-99B Histological analysis was performed by conventional light microscopy.

[0042] The experimental results of this example are as figure 2 (400 times) as shown:

[0043] At each time point, DRG neurons from the normal group, the sham group, the nerve root exposure group, and the 4-day nerve root compression group presented large, round shapes with intact structures. The cytoplasm of each group showed prominent Nissl bodies and the nuclei were slightly stained. Neurons are surrounded by satellite cells and arranged regularly;

[0044] 10 days after nerve root compression injury, DRG neurons of the surgically operated group exhibited irregular arrangements and vacuoles formed in their cytoplasm.

[0045] When the compres...

Embodiment 3

[0047] The DRG slices were stained with TUNEL reagent, and the TUNEL staining in this example used an in situ cell death detection kit (MBL, 8445, Japan). Specifically, the DRG slices were washed with PBS (twice, 5 min each), and incubated in 50 μL TdT buffer II at room temperature for 10 min. After removing TdT buffer II, the DRG slices were incubated in 50 μL TdT solution at 37°C for 60 min, and then soaked in TB solution and incubated at room temperature for 15 min. Nuclei were counterstained with 1 μg / mL DAPI (BioChemica). After labeling, the stained DRG sections were washed with PBS. Analysis was then performed using an immunofluorescence microscope (Olympus).

[0048] The experimental results of this example are as image 3 Shown:

[0049] In order to confirm the occurrence of apoptosis, this example uses TUNEL analysis to detect DNA strand breaks. image 3 In situ TUNEL staining of DRGs from control and operated animals is shown ( image 3 A, 400 times) and the pe...

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Abstract

The invention discloses a construction method of an animal model of a rat with compressive injury of nerve roots. The construction method comprises the following steps: (1) preparing a right-angled triangular rubber block; (2) completely exposing the nerve root L5 of the rat through an operation; (3) inserting the rubber block into the spinal canal and pressing the nerve root L5; (4) sewing the cuts layer by layer to fix the rubber block until the rubber block cannot be inserted; (5) normally feeding rats, regularly observing histological change conditions of dorsal root ganglion and apoptosisconditions of dorsal root ganglion of the rat, detecting activity conditions of caspase-3, and observing protein expression conditions of cytokines. According to the construction method, the histopathology, cell apoptosis condition, activity condition of caspase-3 and expression condition of various cell factors of the animal model of the rat with compressive injury of nerve roots constructed bythe method are all similar to pathological characteristics of clinical spinal degenerative diseases.

Description

technical field [0001] The invention belongs to the technical field of medical animal model construction, and in particular relates to a method for constructing a rat animal model of nerve root compression injury. Background technique [0002] Degenerative conditions of the spine, including herniated discs, ossification of the paravertebral ligaments, and hypertrophy of the facet joints, can lead to nerve root compression and subsequent abnormalities in nerve conduction. It has been studied extensively for many years, but its pathological mechanism remains unclear. Recently, it has been reported that sciatic nerve injury and nucleus pulposus herniation induces apoptosis of some types of cells in the dorsal root ganglion (DRG). Previous studies have demonstrated that neuropathic pain is dependent on DRG cell death following nerve injury. In addition, previous studies have also demonstrated that nerve root compression injury induces apoptosis of DRG neurons. Although many p...

Claims

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Application Information

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IPC IPC(8): A01K67/027A61K49/00
CPCA01K67/027A01K2207/30A01K2227/105A01K2267/0318A61K49/0008
Inventor 徐乐勤周瑞娟卢晓惠陈海鹏林桦楠
Owner 厦门市中医院
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