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Construction and evaluation methods of mouse ischemic stroke model

A technology for ischemic stroke and mice, applied in the field of biomedicine, can solve the problems of difficult clinical transformation and needs to be improved, and achieve the effect of close pathological mechanism, less trauma and high postoperative survival rate

Inactive Publication Date: 2021-08-13
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the problem of this method in reconstructing the ischemic state of brain tissue is that PTS not only causes cytotoxic edema of brain tissue, but also causes cerebral angioedema. In clinical practice, cytotoxic edema mainly occurs when human ischemic stroke occurs, and the blood-brain barrier is weak. It will be damaged immediately, which is also a major reason for the difficulty of clinical transformation
[0005] At present, the clinical prevention and treatment strategy for acute stroke is mainly intravenous injection of tissue plasminogen activator within 4.5 hours after stroke, but its thrombolytic efficiency and safety still need to be improved, and it is important for early intervention of stroke and later neurological function. There are still many problems in the recovery

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  • Construction and evaluation methods of mouse ischemic stroke model
  • Construction and evaluation methods of mouse ischemic stroke model
  • Construction and evaluation methods of mouse ischemic stroke model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Amphiphilic, positively charged near-infrared fluorescent molecules can be used as probes for thrombus in stroke patients

[0029] Thrombectomy was performed in patients with acute ischemic stroke, and the fresh thrombus was stored at -80°C. Frozen tissue samples were cut into 10 μm thin slices using a cryostat, pasted on a clean slide, and stored in a slide box at -80°C. The frozen tissue sections were taken out from -80°C and placed at room temperature for 10 min. After the sections returned to room temperature and dried, the OCT was washed with PBS and then stained. The fluorescent molecule ZnPc(Lys) 5 (3.5 μM) was added dropwise on tissue sections, incubated at room temperature for 2 h, and washed 3 times with PBS. After ZnPc(Lys) 5 The stained and labeled thrombus section samples were imaged under a laser confocal microscope with an excitation wavelength of 633 nm and a magnification of 10×.

[0030] see results figure 1 B, The results show that the t...

Embodiment 2

[0031] Example 2: Construction of a mouse model of ischemic stroke

[0032] After the mice were successfully anesthetized, they were fixed on the operating table in the supine position. The skin was prepared and disinfected routinely. A midline neck incision was made, and the glandular tissue and fascia of the neck were bluntly dissected, and the CCA and ECA were exposed and separated. The ECA was ligated at the bifurcation of the CCA, ECA, and ICA, and a thread was embedded at the proximal end of the CCA. The fluorescent probe (50 nmol / kg) was injected into the blood circulation through the mouse tail vein. After 5 minutes, the proximal end of the CCA was ligated, and the exposed CCA length between the two ligations should be about 13 mm. Hook the exposed CCA into the groove of the thrombus probe of the YLS-14B thrombogenesis instrument, gently lower the pressure plate, keep the thrombus probe perpendicular to the body of the mouse, fix the upper end of the probe on the brac...

Embodiment 3

[0034] Example 3: Evaluation of a mouse model of ischemic stroke using a small animal in vivo imager

[0035] After the operation, the mice were depilated from the skin of the brain and placed in a small animal live imager (FMT 2500 TM Real-time monitoring of ZnPc(Lys) in mouse brain in LXinstrument, PerkinElmer) 5 fluorescent signal. The instrument uses a 680nm laser diode to excite ZnPc(Lys) 5 Molecule, select the mouse brain as ROIs, scan 50-60 source positions (the distance between adjacent scanning points is 3 mm), and obtain the localization results of thrombus. At the same time, mice in the sham-operated group were used as controls.

[0036] see results image 3 , the brain of the affected side of the mice in the model group showed strong fluorescence, and the fluorescence intensity did not decrease significantly after 24 hours, indicating that the thrombus formed by direct current stimulation was relatively stable and could stably block the cerebral artery on the ...

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Abstract

The invention provides a construction method of a mouse ischemic stroke model. According to the construction method, current stimulation is given to a Common carotid artery (CCA) of a mouse to form a thrombus embolus, the thrombus embolus is activated, the thrombus embolus is made to spontaneously enter the cranium through an Internal carotid artery (ICA), the intracranial Middle artery (MCA) is blocked, and then acute brain blood supply blocking and brain dysfunction are caused. The invention also provides an evaluation method of the mouse ischemic stroke model. The evaluation method comprises the following steps: marking the thrombus embolus formed on the CCA by using a near-infrared fluorescent molecular probe, tracking the position of the thrombus embolus in the mouse cranium by using a small animal living imaging instrument, and evaluating the ICA clogging degree by using fluorescence intensity.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for constructing and evaluating a mouse ischemic stroke model. Background technique [0002] Stroke, also known as cerebrovascular accident (CVA), refers to the stenosis, occlusion or rupture of cerebral arteries caused by various factors, resulting in acute cerebral blood circulation disorder and brain blood supply blockage, which in turn leads to cranial nerve function. obstacle. Stroke is the second leading cause of human death, second only to ischemic heart disease, accounting for 9% of global human death. my country is a country with a high incidence of stroke, and the incidence of stroke is as high as 1%. The "China Stroke Prevention Report 2018" shows that from 2016 to 2018, the incidence of ischemic stroke in my country still showed a continuous upward trend. The symptoms of stroke are related to the damaged part, so the symptoms are diverse, and the main features ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027A61B5/026
CPCA01K67/027A61B5/0261A01K2207/30A01K2227/105A01K2267/03
Inventor 黄明东陈丹袁彩江龙光徐芃王志友刘沛文
Owner FUZHOU UNIV
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