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Preparation method and application for Parkinson's disease rat model

A Parkinson's disease and rat model technology, applied in the field of biology, can solve the problems of restricting the treatment method of PD pathogenesis, and greatly affecting the success rate of PD modeling, so as to achieve increased range of action, high success rate, and simple operation Effect

Inactive Publication Date: 2015-10-28
SHANDONG NORMAL UNIV
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Problems solved by technology

In many previous reports, one or two sites in the substantia nigra pars compacta, medial forebrain bundle, striatum, and ventral dorsal tegmental area of ​​the midbrain were selected for PD modeling. The selection of different sites in the tract, striatum, and ventral tegmental region of the midbrain has a great influence on the success rate of PD modeling. The success rate of PD modeling reported so far is mostly below 80%, which seriously restricts the PD modeling. Research on pathogenesis and treatment methods

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  • Preparation method and application for Parkinson's disease rat model
  • Preparation method and application for Parkinson's disease rat model

Examples

Experimental program
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Embodiment 1

[0020] Embodiment 1: Preparation of Parkinson's disease rat model

[0021] After intraperitoneally injecting 4% chloral hydrate (400mg / kg) to anesthetize the rats, fix the rat's head on the brain stereotaxic instrument, cut off the hair on the top of the skull, and after routine disinfection, cut the skin on the top of the skull along the midline. , Posterior fontanel, refer to the sixth edition of the brain stereotaxic atlas of "The Rat Brain in Stereotatic Coordinates" edited by Paxinos G et al. Site, coordinates: 1.72mm behind bregma, 2.13mm lateral to the midline, inject 1.5ul of 6-OHDA at 8.5mm and 8.7mm below the surface of the skull, and inject the needle slowly at a speed of 1mm / min. , the injection speed is 1ul / min, and the needle is retained for 10 minutes after the injection is completed. 80,000 units of penicillin were injected intraperitoneally for 3 consecutive days to prevent infection. Two groups of experimental animals were injected with apomorphine hydrochl...

Embodiment 2

[0024] After 4 weeks of successful modeling in Example 1, the rats were overdose anesthetized with chloral hydrate, the thorax was opened, the perfusion needle was inserted into the aorta from the left ventricle for perfusion, and 250 ml of 0.01M PBS solution was first poured into the body to flush out the blood in the body , and then filled with 250ml of 4% paraformaldehyde solution for tissue fixation. After the perfusion, use bone forceps to crush the skull and peel off the brain tissue, put the brain tissue in the same fixative solution for 24 hours, put it in 30% sucrose solution for dehydration until the brain tissue sinks to the bottom, and then take it out in the freezer Freeze and cut out substantia nigra brain slices in a microtome with a thickness of 25um, take one every 4 slices, wash the brain slices with PBS and wash them with 3% H 2 o 2 React with methanol solution for 30 minutes, then block with goat serum overnight, add rabbit anti-tyrosine hydroxylase antibo...

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Abstract

The invention discloses a preparation method for a Parkinson's disease rat model. The preparation method comprises the following steps: obtaining a locus of a rat MFB according to a brain three-dimensional positioning map or / and magnetic resonance imaging data of an experimental rat, wherein the locus is in the position which is 1.72 mm behind the bregma and 2.13 mm beside a middle line; respectively injecting 1-2 [um]l of 6-OHDA in positions 8.5 mm and 8.7 mm below the surface of the skull; after model making, injecting apomorphine hydrochloride to modeled animals for four weeks continuously for carrying out rotary experimental detection, and rotating towards the normal side, if the number of turns is larger than 7 r / min, regarding that model making is successful, and if the rotating direction is wrong or the number of revolutions is not enough, regarding that model making is unsuccessful. According to the invention, medicine is injected and fed to the relatively thick part of the MFB, and 6-OHDA damages nigral dopaminergic neurons through the dopaminergic neuron axon retrograde property, so that the success rate of model making is high, the success rate of 84% can be realized one week after model making and the success rate can reach 92% two weeks after model making; for model making reported at present, model making at other selected loca can be successful until four weeks mostly and the success rate of model making is usually lower than 80%.

Description

technical field [0001] The present invention relates to the field of biology, in particular, the present invention relates to a preparation method and application of a Parkinson's disease rat model. Background technique [0002] Parkinson's disease (PD) is the most common neurodegenerative disease after Alzheimer's disease, with degeneration and loss of DA neurons in the substantia nigra pars compacta as the main pathological change. The incidence rate in the elderly population accounts for about 0.5%-1%, and the incidence rate can reach 1%-3% in the elderly over 80 years old. Although in recent years, with the in-depth research on Parkinson's disease, some treatment methods have been able to alleviate some symptoms of the disease, but none of them can achieve the purpose of complete cure. Therefore, more in-depth research on the pathogenesis and treatment of PD Methods are necessary, and a stable and efficient animal model is a prerequisite for in-depth research on the dis...

Claims

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Application Information

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IPC IPC(8): A61K31/137A61K31/473A61K49/00
Inventor 王敏谢金鹿何婷婷李敏耿希文祝建平张晓曲庆洋类成东侯亚兵杨茂全
Owner SHANDONG NORMAL UNIV
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